Ly-6B.2 Alloantigen antibody | 7/4
Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by immunofluorescence.
Image caption:
CX3CR1+, Ly6Clo monocytes are present in perihematomal brain in CX3CR1-null BM chimeras. Mice were injected intracardially with 150 ug tomato lectin to delineate vasculature. Brain slices were stained with Ly6B.2 antibody to differentiate Ly6Chi monocytes and neutrophils from Ly6Clo CX3CR1+ monocytes and Dapi nuclear stain. A) Intracardiac injection of lectin stains vasculature in the brain– 20x. B) At 72 hours after ICH, distinct subsets of amoeboid Ly6B.2+ (Ly6Chi monocytes or neutrophils) and GFP+ (CX3CR1+) monocytes are seen in the perihematomal region. 20x. C) At 7 days after ICH, amoeboid, Ly6B.2−,CX3CR1+ cells are seen in the brain parenchyma surrounding the ICH cavity. The center of the ICH cavity is filled with Ly6B.2+ (Ly6Chi) monocytes or neutrophils. Tomato lectin staining is also seen in the center of the ICH cavity, most likely due to blood brain barrier breakdown at this time point leading to nonspecific lectin staining of myeloid cells. 10x. White box indicates the inset shown in panel E. D) At 14 days, amoeboid, CX3CR1+ cells can still be seen in the perihematomal region, however more CX3CR1+ cells have a ramified morphology. No Ly6B.2+ (Ly6Chi monocytes or neutrophils) cells are found. Overall blood-brain barrier disruption is improved leading to less lectin staining in the cavity, although there remain some lectin+ cells (CX3CR1+ monocytes and CX3CR1− microglia). 10x. White box indicates inset shown in panel F. E) 20x image of panel C – perihematomal region 7 days after ICH F) 20x image of panel D – perihematomal region 14 days after ICH. Blue – Dapi, GFP – CX3CR1, Red – Tomato lectin, Pink – Ly6B.2, n = 5.
From: Taylor RA, Hammond MD, Ai Y, Sansing LH (2014)
CX3CR1 Signaling on Monocytes Is Dispensable after Intracerebral Hemorrhage.
PLoS ONE 9(12): e114472.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 antibody, clone 7/4 used for the detection of Ly-6B.2 positive cells by immunohistochemistry.
Image caption:
Effect of atorvastatin on platelet and neutrophil infiltration after reperfusion. The infiltration of platelets (upper panel) and neutrophils (lower panel) were evaluated by immunohistochemistry in the 4 chimeric groups that received 40 min of LAD occlusion and 60 min of reperfusion. Platelets and neutrophils were found predominantly in the ischemic area in B6/B6 chimeras. Atorvastatin reduced both platelet and neutrophil infiltration in B6/B6 and B6/KO chimeras, but not in KO/B6 chimeras.
From: Tian Y, Linden J, French BA, Yang Z (2014)
Atorvastatin at Reperfusion Reduces Myocardial Infarct Size in Mice by Activating eNOS in Bone Marrow-Derived Cells.
PLoS ONE 9(12): e114375.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by immunohistochemistry.
Image caption:
Post-ischemic treatment with HDL does not decrease leukocyte infiltration. Mice were submitted to LAD occlusion for 45min and hearts were reperfused for 24h. Mice were injected or not (control mice IR) with native HDL (nHDL), rHDLB (apoAI + POPC + S1P) one minute before reperfusion. A. Quantification of infiltrated neutrophils (Ly-6B.2+ cells) per area in frozen sections of infarcted hearts at 24h of reperfusion. B. Representative images of neutrophil (Ly-6B.2+ cells) infiltration. C. Quantification of infiltrated neutrophils (Ly6G+ cells) per area in frozen sections of infarcted hearts at 24h of reperfusion. D. Representative images of neutrophil (Ly6G+ cell) infiltration. E. Quantification of infiltrated macrophages (CD68+ cells) per area in frozen sections of infarcted hearts at 24h of reperfusion. F. Representative images of macrophage (CD68+ cells) infiltration. Data are expressed as scattered plots (mean±SEM, n = 7–10 per group). Results are expressed as percentages of stained area on total heart surface area. No significance difference between groups was found using unpaired-student t-test.
From: Brulhart-Meynet M-C, Braunersreuther V, Brinck J, Montecucco F, Prost J-C, et al. (2015) Improving Reconstituted HDL Composition for Efficient Post-Ischemic Reduction of Ischemia Reperfusion Injury. PLoS ONE 10(3): e0119664.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of neutrophils in mouse abdominal aorta by immunogluorescence.
Image caption:
Flow cytometric analysis of bead-labelled aortic leukocytes in ApoE−/− and Rgs1−/− ApoE−/− mice that received Ang II infusion at 0.8 mg kg−1 per day for 5 days following fluorescent bead labelling of circulating inflammatory monocytes. (a) Representative dot plots shown for gated aortic cells of each positive population from ApoE−/− and Rgs1−/− ApoE−/− mice with representative percentages. Labels on both axes are on a log scale. (b) Quantification of the number of bead-labelled CD45+ cells in aortas of Ang II-infused mice at days 3 and 5. (c) Immunofluorescence microscopy of abdominal aortas from ApoE−/− mice at day 5 after Ang II infusion and bead labelling stained for Ly6C and 7/4 (red), 4′,6-diamidino-2-phenylindole (blue). Arrows indicate the presence of cells containing fluorescent beads (green) on the luminal side (L) of the aorta or within the internal elastic laminar (green autofluorescence). Adv, adventitia. **P<0.01 calculated using the Student’s t-test (n=5–7 per group. Data in b are expressed as mean±s.e.m.).
From: Patel J, McNeill E, Douglas G, Hale AB, de Bono J, Lee R, Iqbal AJ, Regan-Komito D, Stylianou E, Greaves DR, Channon KM.
RGS1 regulates myeloid cell accumulation in atherosclerosis and aortic aneurysm rupture through altered chemokine signalling.
Nat Commun. 2015 Mar 18;6:6614.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of infiltrating neutrophils in a murine colitis model by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
Nur77-/- mice develop augmented DSS-induced colitis. The body weight of the mice was determined daily during DSS treatment (A). Upon harvest colon weight, spleen weight (B) and colon length (C) were determined. The disease activity index (DAI) was calculated as an average in weight loss, stool consistency and bleeding scores (D). Colon histology was scored on hematoxylin-eosin stained sections (E) for the parameters shown (F). Neutrophil numbers were counted in healthy and DSS colon sections and the influx was calculated by taking the ratio of Healthy / DSS (G). Representative photomicrographs are shown with original magnification 100x. Colon pieces were cultured overnight and cytokine levels were determined in the supernatant and corrected for mg of tissue (H). Data are presented as mean±SEM; *p<0.05, **p<0.01, ***p<0.001. Scale bars represent 1mm. ME = Muscularis externa, KC = Chemokine ligand 1 (mouse homologue of IL-8; a potent neutrophil attractant).
From: Hamers AAJ, van Dam L, Teixeira Duarte JM, Vos M, Marinković G, van Tiel CM, et al. (2015)
Deficiency of Nuclear Receptor Nur77 Aggravates Mouse Experimental Colitis by Increased NFκB Activity in Macrophages.
PLoS ONE 10(8): e0133598.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
Increased Inflammatory Cell Infiltration and Inflammatory Mediators in Placental Tissues Following P.g. Infection. Immunohistochemical staining of gd 15 placental tissues of NC and P.g.-infected groups (n = 6 for each group). (A,B) PMNLs, (C,D) F4/80 positive macrophages, (E,F) TNF-α positive macrophages, (G, H) COX-2 positive staining, (I, J) CD-31 stained endothelial cells, (K,L) 8OHdG (oxidative DNA stress marker), (M,N) cleaved-caspase 3 (apoptosis marker), and (O,P) histomorphometric analysis of PMNLs and F4/80 (macrophages). Scale bars, 100μm. Statistical significance was determined using the Student t-test. *p<0.05, **p<0.01. Olympus BH2 microscope and Nikon digital sight DS-L2 camera were used for capturing images. (Q) Representative results of mRNA expression of COX-2, Gal-3, IL-8 and TNF-α in placental tissues in NC group and P.g.-infected group (n = 3 for each), GAPDH was used as internal control. Molecular weights were labeled to the right of each band. Yellow dashed square shows that PMNLs are restricted to the blood vessels.
From: Ao M, Miyauchi M, Furusho H, Inubushi T, Kitagawa M, Nagasaki A, et al. (2015)
Dental Infection of Porphyromonas gingivalis Induces Preterm Birth in Mice.
PLoS ONE 10(8): e0137249.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Ly-6B.2 Alloantigen antibody, clone 7/4 (MCA771GA) used for the evaluation of Ly6B.2 expression on various leukocyte types by flow cytometry.
Image caption:
A) First, CD45+ cells were selected based on a FSC-A/CD45 profile followed by gating on single cells (SSC-A/FSC-W profile). Then, CD11b+ cells were selected using an CD11b/FSC-A profile within the CD45+ population. Subsequently, neutrophils (CD11b+Ly6cintLy6G+) were identified using a Ly6G/FSC-A profile and the remaining cells were used in an Ly6C versus MHC-II profile to identify monocytes (CD11b+Ly6chighLy6G-MHC-II-), monocyte-derived macrophages (CD11b+Ly6chighLy6G-MHC-II+), resident macrophages (CD11b+Ly6c-Ly6G-MHC-II-) and a Rest fraction (CD11b+Ly6c-Ly6G-MHC-II-). B) Surface marker expression on different myeloid cell subsets. Expression of F4/80, CCR2, Ly6B, MerTK and CD64 is displayed. C. Myeloid cell composition of liver in absolute cell numbers. D. Myeloid cell composition of spleen in absolute cell numbers. Values represent mean ± SD of 4 mice per group. *: p-value < 0.05 and if nothing is mentioned the differences were not significant.
From: Citation: Cnops J, De Trez C, Stijlemans B, Keirsse J, Kauffmann F, Barkhuizen M, et al. (2015)
NK-, NKT- and CD8-Derived IFNγ Drives Myeloid Cell Activation and Erythrophagocytosis, Resulting in Trypanosomosis-Associated Acute Anemia.
PLoS Pathog 11(6): e1004964.
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Rat anti Mouse Ly-6B.2 monoclonal antibody, clone 7/4 (MCA771GA) used for the detection of inflammatory neutrophils in mouse lung tissue sections by immunofluorescence
Image caption:
Apigenin reduces LPS-induced apoptosis in the lungs. Lungs from mice treated with LPS, control, apigenin (Api) or apigenin 3 h prior to LPS (Api + LPS) were obtained at 24 h. (A) The number of apoptotic cells was determined by the TUNEL assay and expressed as the mean ± SEM of the numbers of stained cells per 1000× microscopic field (cpf) (N = 4 mice for each biological condition; * p < 0.01 vs. LPS), scale bars: 25 μm for all figures; (B,C) Caspase-3 activity was determined by the DEVD-AFC assay using lung lysates as described in materials and methods. Values represent the means ± SEM (N = 4 mice for each biological condition; * p < 0.05 vs. LPS); (D) Lung tissue immuno-histochemistry using antibodies specific for active caspase-3 in combination with anti-CD31, anti-cytokeratin or anti-7/4 antibodies specific for endothelia, epithelia and neutrophils, respectively. Scale bars, 100 μm.
From: Cardenas, H.; Arango, D.; Nicholas, C.; Duarte, S.; Nuovo, G.J.; He, W.; Voss, O.H.; Gonzalez-Mejia, M.E.; Guttridge, D.C.; Grotewold, E.; Doseff, A.I.
Dietary Apigenin Exerts Immune-Regulatory Activity in Vivo by Reducing NF-κB Activity, Halting Leukocyte Infiltration and Restoring Normal Metabolic Function.
Int. J. Mol. Sci. 2016, 17, 323.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 monoclonal antibody, clone 7/4 (MCA771GA) used for the identification of infiltrating neutrophils in murine pulmonary tissue sections by immunohistochemistry.
Image caption:
Apigenin decreases LPS-induced neutrophil infiltration in the lungs. Lungs from mice receiving Api + LPS, LPS, Api or vehicle (control) were collected at 24 h post-challenge. (A) H&E staining; (B) Chloroacetate esterase (CAE) staining; (C) Lungs stained with anti-7/4 antibodies; (D) Leukocyte infiltration determined by counting CAE-positive cells per high power field (cpf) in lungs used in (B); (E) Number of infiltrated neutrophils as determined by counting stained cells per cpf in in lungs used in (C). Data represent the mean ± SEM (N = 9 mice for each biological condition; * p < 0.05 vs. LPS). Scale bars for all figures are 25 μm and 100 μm for insets.
From: Cardenas, H.; Arango, D.; Nicholas, C.; Duarte, S.; Nuovo, G.J.; He, W.; Voss, O.H.; Gonzalez-Mejia, M.E.; Guttridge, D.C.; Grotewold, E.; Doseff, A.I.
Dietary Apigenin Exerts Immune-Regulatory Activity in Vivo by Reducing NF-κB Activity, Halting Leukocyte Infiltration and Restoring Normal Metabolic Function.
Int. J. Mol. Sci. 2016, 17, 323.
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Rat anti Mouse Ly-6B.2 monoclonal antibody, clone 7/4 (MCA771G) used for the detection of neutrophils in murine kidney tissue sections by immunohistochemistry. Macrophages were detected using Rat anti Mouse F4/80 antibody, clone A3-1 (MCA497 )
Image caption:
Immunostaining of macrophages and neutrophils in the kidneys of M-Rac1 FC and KO mice. (A) Representative micrographs of F4/80 immunostaining in the kidneys of Vehicle- or LPS-injected M-Rac1 FC and KO mice. Original magnification x 200. (B) Quantitative analysis of F4/80-positive macrophages. Data are means ± s.e.m. Statistical analysis was performed by two-way ANOVA, n.s. genotype effect, P < 0.01 treatment effect, n.s. interaction effect. **P < 0.01 by Bonferroni's post hoc test. n = 5 per each group. (C) Representative micrographs of Ly-6B.2 (mAb 7/4) immunostaining. (D) Quantitative analysis of Ly-6B.2-positive neutrophils. Statistical analysis was performed by two-way ANOVA, n.s. genotype effect, P < 0.01 treatment effect, n.s. interaction effect. **P < 0.01 by Bonferroni's post hoc test. n = 6 per group.
From: Nagase M, Kurihara H, Aiba A, Young MJ, Sakai T (2016)
Deletion of Rac1GTPase in the Myeloid Lineage Protects against Inflammation-Mediated Kidney Injury in Mice.
PLoS ONE 11(3): e0150886.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 monoclonal antibody, clone 7/4 (MCA771GA) used to identify neutrophils in formalin fixed, paraffin embedded murine placental tissue sections.
Image caption:
Increased Inflammatory Cell Infiltration and Inflammatory Mediators in Placental Tissues Following P.g. Infection.Immunohistochemical staining of gd 15 placental tissues of NC and P.g.-infected groups (n = 6 for each group). (A,B) PMNLs, (C,D) F4/80 positive macrophages, (E,F) TNF-α positive macrophages, (G, H) COX-2 positive staining, (I, J) CD-31 stained endothelial cells, (K,L) 8OHdG (oxidative DNA stress marker), (M,N) cleaved-caspase 3 (apoptosis marker), and (O,P) histomorphometric analysis of PMNLs and F4/80 (macrophages). Scale bars, 100μm. Statistical significance was determined using the Student t-test. *p 7lt; 0.05, **p < 0.01. Olympus BH2 microscope and Nikon digital sight DS-L2 camera were used for capturing images. (Q) Representative results of mRNA expression of COX-2, Gal-3, IL-8 and TNF-α in placental tissues in NC group and P.g.-infected group (n = 3 for each), GAPDH was used as internal control. Molecular weights were labeled to the right of each band. Yellow dashed square shows that PMNLs are restricted to the blood vessels.
From: Ao M, Miyauchi M, Furusho H, Inubushi T, Kitagawa M, Nagasaki A, et al. (2015)
Dental Infection of Porphyromonas gingivalis Induces Preterm Birth in Mice.
PLoS ONE 10(8): e0137249.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the identification of neutrophils in the distal colonic mucosa by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
Increased T cell and neutrophil infiltration in the colonic mucosa of DKO/AT mice is accompanied by altered MLN Treg phenotype.
(A and B) Representative images of immunohistochemical analysis of (A) CD4+ cell abundance and (B) neutrophil abundance in the distal colonic mucosa; (C) qPCR analysis of mucosal expression of neutrophil collagenase MMP8; (D) Flow cytometry analysis of regulatory T cells in mesenteric lymph nodes (MLN) in adoptively transferred mice and effect of antibiotics. Bar graphs represent mean with SD values. Statistical analysis were performed using one-way ANOVA (P value indicated in each panel) with Fisher's LSD post-hoc test. Asterisks in all panels indicate statistical significance: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
From: Laubitz D, Harrison CA, Midura-Kiela MT, Ramalingam R, Larmonier CB, Chase JH, et al. (2016)
Reduced Epithelial Na+/H+ Exchange Drives Gut Microbial Dysbiosis and Promotes Inflammatory Response in T Cell-Mediated Murine Colitis.
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Rat anti Mouse LY-6B.2 antibody, clone 7/4 (MCA771G) used for detection of neutrophils in murine skin implants by immunohistochemistry.
Image caption:
Sample d-TT (A) and d-TT EW (B) were implanted subcutaneously for 3 days before excision and immunohistochemical analysis. d-TT showed increased Ly-6B-positive cells (neutrophils) compared to d-TT EW (scale bar is 50 μm). Quantification of staining (C) shows increases in Ly-6B in d-TT. *p <0.05.
From: Morris Aaron H., Chang Julie, and Kyriakides Themis R.
BioResearch Open Access. July 2016, 5(1): 177-87.
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Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771G) used for the identification of neutrophils in an alchohol induced model of liver injury by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
RIP3 KO mice decrease the hepatic expression of inflammatory cytokine genes but have no effects on Gao-binge alcohol-induced neutrophil infiltration
WT and RIP3 KO mice were treated with the Gao-binge model. A. RNA from mouse livers was used to measure gene expression by qPCR. Results were normalized to β-actin and expressed as fold change compared to WT control. Data shown are means ± S.E (n = 3-4 mice per group. *p<0.05; by one-way ANOVA). B. Representative images were shown from Ly6B immunohistochemistry after Gao-binge treatment. Arrows denote the Ly6B positive neutrophils. C. The number of Ly6B positive neutrophils was counted from at least 22 areas (400 X magnification) each mouse. Results shown are means ± S.E. (n = 4-6 mice per group. **p<0.01 compared to WT control by one-way ANOVA).
From: Wang S, Ni HM, Dorko K, Kumer SC, Schmitt TM, Nawabi A, Komatsu M, Huang H, Ding WX.
Increased hepatic receptor interacting protein kinase 3 expression due to impaired proteasomal functions contributes to alcohol-induced steatosis and liver injury.
Oncotarget. 2016 Apr 5;7(14):17681-98.
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Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by immunofluorescence.
Image caption:
Confocal neutrophil-cytokine double labeling experiments using incised skin. For these studies skin from the wound edges was harvested 2 hours after incision. After sectioning, the tissue was exposed to anti-neutrophil and anti-IL-1β antibodies followed by the application of CY3 (green fluorescence, neutrophils, panel A) and FITC (red, IL-1β, panel B) conjugated secondary antibodies. Panel C presents the merged image with arrows pointing to several strongly double labeling cells. The scale bar in panel C is 50 μm in length. These micrographs were taken of dermal tissue under 630× magnification.
From: Clark JD, Shi X, Li X, Qiao Y, Liang D, Angst MS, Yeomans DC. Morphine reduces local cytokine expression and neutrophil infiltration after incision. Mol Pain. 2007 Oct 2;3:28
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Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by flow cytometry.
Image caption:
Characterization of high dose zymosan peritonitis. A) Representative flow-cytometric density plots showing the recruitment of inflammatory cells and the clearance of fluorescent (FITC)-labelled zymosan particles. The examples shown are taken from 18 hours after the administration of zymosan. Cells were identified as described in the methods. The panel on the left shows the gating of Ly-6B+ cells, which includes (right) Ly-6Ghigh neutrophils and Ly-6G− monocytes and Mø. Both neutrophils and monocytes/Mø exhibit association with FITC-labeled zymosan at this time point (right panel). B) Graphical representation of the number of neutrophils (left) and all types of monocyte (Mo) and Mø combined (right) in the peritoneal cavity before and after acute zymosan peritonitis. n = 3 129S6/SvEv mice per group and is representative of 2 independent experiments. Data is shown as mean ± SEM and cells were isolated from naïve animals (0 hours), and challenged mice 4, 18, 72 and 168 hours after induction of peritonitis. C) Graphical representation of the percentage of neutrophils and Mo/Mø present that are associated with zymosan, scored by flow-cytometry as indicated in (A) above. D) Photomicrograph, which is representative of neutrophils associated with zymosan in a 4 hour inflammatory infiltrate. Those neutrophils that are associated tend to have multiple particles, but only represent a minority of the total neutrophils present at this time (see (C) above).
From: McDonald JU, Rosas M, Brown GD, Jones SA, Taylor PR (2012)
Differential Dependencies of Monocytes and Neutrophils on Dectin-1, Dectin-2 and Complement for the Recognition of Fungal Particles in Inflammation.
PLoS ONE 7(9): e45781.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by immunohistochemistry.
Image caption:
Peri-incisional infiltration of neutrophils, immunohistochemical appearance. Hind paw skin from incised mice was processed for the identification of neutrophils. Panel A displays an H&E stained section of plantar hind paw skin demonstrating a predominantly dermal cellular infiltrate 2 hours after incision. The dermal and epidermal layers are labeled. Panel B displays the appearance of non-incised hind paw skin stained with a neutrophil-specific antibody. Panel C displays a micrograph taken of incised skin stained for neutrophils 2 hours after incision. Note the abundance of darkly staining infiltrating neutrophils in the dermis of this section compared to that displayed in panel B. The scale bar in panel C is 150 μm in length. All micrographs were taken using 200× magnification.
From: Clark JD, Shi X, Li X, Qiao Y, Liang D, Angst MS, Yeomans DC. Morphine reduces local cytokine expression and neutrophil infiltration after incision. Mol Pain. 2007 Oct 2;3:28.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by immunohistochemistry.
Image caption:
Renal inflammatory changes in Ntn1+/− mice following ischemia. Ntn1+/− mice and their respective age-, weight-, and gender-matched littermate controls (Ntn1+/+) were subjected to 30 minutes of left renal artery ischemia. (A–D) Neutrophil staining. Arrows indicate neutrophils (magnification 400×). (E) Quantification of neutrophil tissue accumulation by measurement of myeloperoxidase (MPO). (F) TNF-α and (G) interleukin-6 (IL-6) and (H) interleukin-10 (IL-10) were assessed by real-time RT-PCR from renal tissues. Data were calculated relative to β-actin and are expressed as fold change compared to sham-operated animals without ischemia (−I). Data are representative of four to six independent experiments for each experimental condition (mean ± SD).
From: Grenz A, Dalton JH, Bauerle JD, Badulak A, Ridyard D, et al. (2011)
Partial Netrin-1 Deficiency Aggravates Acute Kidney Injury. PLoS ONE 6(5): e14812.
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Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by immunofluorescence.
Image caption:
Recruitment of inflammatory cells in the intestine of neonates after CpG-ODN treatment. Eight-day-old neonatal mice received 20 μg/g of CpG-ODN orally. Twenty-four hours after treatment, ilea were removed to analyze the recruitment of inflammatory cells by immunohistology. Six to ten neonates from different litters (5 sections for each animal) per group were analyzed. Data were analyzed by non-parametric Mann-Whitney test.
From: Lacroix-Lamandé S, Rochereau N, Mancassola R, Barrier M, Clauzon A, et al. (2009)
Neonate Intestinal Immune Response to CpG Oligodeoxynucleotide Stimulation.
PLoS ONE 4(12): e8291.
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Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by immunofluorescence.
Image caption:
166 mM H2O2 increased neutrophil infiltration in day 1 and 6 wounds. Fluorescence intensity of the neodermis was quantified using ImageJ. The area quantified is outlined with the dashed line. Results shown are mean ± S.E.M, n = 6-7. A representative section from each treatment is shown. ES – Eschar; HE – Hyper-proliferating epidermis; ND – neodermis.*p<0.05.
From: Loo AEK, Wong YT, Ho R, Wasser M, Du T, et al. (2012) Effects of Hydrogen Peroxide on Wound Healing in Mice in Relation to Oxidative Damage.
PLoS ONE 7(11): e49215
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Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by flow cytometry.
Image caption:
F4/80 and CD11b+ cells infiltrating the iris after the intracameral injection of antigen also express Ly6C, 7/4 (Ly 6B) and CD45. Preparations of naïve iris cells and cells recovered after an intracameral injection of antigen were stained with anti- F4/80 and (A) Ly6C and 7/4 (B) show a characteristic CD45 peak C) Ly6Glo or negative, CD115, CD49b+, CD62Llo. The figures are representative of 2 experiments. All the isotype controls in the histograms are shown as shaded.
From: Pais R, Bhowmick S, Chattopadhyay S, Lemire Y, Sharafieh R, et al. (2012)
An Intracameral Injection of Antigen Induces In Situ Chemokines and Cytokines Required for the Generation of Circulating Immunoregulatory Monocytes.
PLoS ONE 7(8): e43182.
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Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by immunohistochemistry.
Image caption:
Intact spleen segments from infected animals were fixed in paraformaldehyde, embedded in OCT, sectioned, and stained with various antibodies for the infiltration of macrophages (FA-11), polymorphonuclear neutrophils, and activated monocytes (7/4Ag) and activated macrophages (MHC-II), as described in the text. All data are representative of at least three individual experiments.
From: Plüddemann A, Hoe JC, Makepeace K, Moxon ER, Gordon S (2009)
The Macrophage Scavenger Receptor A Is Host-Protective in Experimental Meningococcal Septicaemia.
PLoS Pathog 5(2): e1000297.
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Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by immunofluorescence.
Image caption:
Neutrophils infiltrate p120 null regions of the small intestine and colon. (A) p120 KO small intestine and colon sections were co-stained for p120 and neutrophils. Arrows and arrowheads indicate regions of p120 ablation. (B) Neutrophils were quantified in oil and tamoxifen-induced small intestine (p = .06) and colon (p = .0092) samples. (C) At the two-month time point, mice were gavaged with FITC-dextran, and serum levels of FITC-dextran were measured to test for an intestinal permeability defect. FITC-dextran levels were significantly higher (3.3-fold) in p120 KO mice (p = .03).
From: Smalley-Freed WG, Efimov A, Short SP, Jia P, Zhao Z, et al. (2011)
Adenoma Formation following Limited Ablation of p120-Catenin in the Mouse Intestine. PLoS ONE 6(5): e19880.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771GA) used for the detection of Ly-6B.2 positive cells by immunohistochemistry.
Image caption:
Changes in markers of hepatic inflammation in wild type and hepatocyte-specific HIF-1β knockout mice after Gao-binge treatment. Age matched male wild type and hepatocyte-specific HIF-1β knockout mice were subjected to Gao-binge treatment. Hepatic mRNA was isolated, and real-time RT-PCR was performed as described in the Materials and Methods (A). Data are presented as means ± SE (n = 3–5). Liver tissue was subjected to immunostaining for neutrophils using an anti-Ly6B antibody and representative images are shown in (B, 40x), and the number of neutrophils were quantified from 10 different fields of each mouse. (C). Data are presented as average number of neutrophils in each filed (means ± SE; n = 4).
From: Ni H-M, Bhakta A, Wang S, Li Z, Manley S, et al. (2014)
Role of Hypoxia Inducing Factor-1β in Alcohol-Induced Autophagy, Steatosis and Liver Injury in Mice. PLoS ONE 9(12): e115849.
This image is from an open access article distributed under the terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 antibody, clone F7/4 (MCA771GA) used for the identifiction of neutrophils in mouse renal sections by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
Honokiol attenuates the increase in plasma creatinine, tubular cell death and neutrophil infiltration induced by renal ischemia-reperfusion (IR) in mice. C57BL/6 mice were subjected to 25 min of bilateral renal ischemia. Honokiol (1 mg/kg) or vehicle was injected twice, 1 h prior to ischemia and 4 h after reperfusion. The blood and kidney tissues were collected at 24 h of reperfusion. (a) Plasma creatinine levels were measured to assess the degree of kidney injury. (b) Representative images of H&E staining of renal sections to evaluate tubular pathology. Arrows indicate necrotic area. (c) Representative images of polymorphonuclear leukocytes (PMN) staining of renal sections to evaluate neutrophil infiltration. (d) Representative images of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining of renal sections to evaluate tubular apoptosis; TUNEL (green) merged with DAPI (blue). Values are expressed as the means ± S.E.M. of 4–8 mice from each group. * p <0.05, ** p <0.01 significant compared with Veh sham. # p <0.05, ## p <0.01 significant compared with Veh IR.
From: Park, E.J.et al.
Honokiol Protects the Kidney from Renal Ischemia and Reperfusion Injury by Upregulating the Glutathione Biosynthetic Enzymes.
Biomedicines 2020, 8, 352.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rat anti Mouse Ly6B.2 antibody, clone F7/4 (MCA771GA) used to identify neutrophils by immunofluorescence.
Image caption:
Heightened wound-induced corneal angiogenesis in Jam-Agt/gt mice.
Confocal images of WT and Jam-Agt/gt eye sections at 0 hours, 3 days, 7 days, and 12 days post full thickness corneal injury. Scale bar 100 μm. Nuclear staining using Draq5 is shown in blue. (C) Ly-6B.2 staining depicts inflammation. (D) Quantitation of Ly-6B.2 staining in the Jam-Agt/gt and WT mice is shown by the ratio of the area covered by Ly-6B.2 to the total corneal area in each image.
From: Chatterjee S, Wang Y, Duncan MK, Naik UP (2013)
Junctional Adhesion Molecule-A Regulates Vascular Endothelial Growth Factor Receptor-2 Signaling-Dependent Mouse Corneal Wound Healing.
PLoS ONE 8(5): e63674.
doi: 10.1371/journal.pone.0063674
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rat anti Mouse Ly6B.2 antibody, clone F7/4 (MCA771GA) used to identify neutrophils by immunofluorescence.
Image caption:
No difference in inflammation induced corneal angiogenesis between Jam-Agt/gt and WT mice.
(A) Photographic images of WT and Jam-Agt/gt eyes. (B) H&E staining of frozen sections prepared from Jam-Agt/gt and WT eyes 12 days post suture. (C&E) Confocal images. Nuclear staining using Draq5 is shown in blue. (C) Corneal inflammation demonstrated by Ly-6B.2 staining. (D) Quantification of Ly-6B.2 staining shows no difference in inflammation between Jam-Agt/gt and WT mice.
From: Chatterjee S, Wang Y, Duncan MK, Naik UP (2013)
Junctional Adhesion Molecule-A Regulates Vascular Endothelial Growth Factor Receptor-2 Signaling-Dependent Mouse Corneal Wound Healing.
PLoS ONE 8(5): e63674.
doi: 10.1371/journal.pone.0063674
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rat anti Mouse Ly6B.2 antibody, clone F7/4 (MCA771GA) used to identify neutrophils by immunofluorescence.
Image caption:
Jam-Agt/gt eyes reveal increased VEGFR-2 signaling during wound-induced corneal angiogenesis.
Confocal images of eye sections of IgG and DC101 treated WT and Jam-Agt/gt mice. Scale bar 100 μm. Nuclear staining using Draq5 is shown in blue. (C) Ly-6B.2 staining showed inflammation in Jam-Agt/gt and WT mice. (D) Quantitation of Ly-6B.2 staining of C.
From: Chatterjee S, Wang Y, Duncan MK, Naik UP (2013)
Junctional Adhesion Molecule-A Regulates Vascular Endothelial Growth Factor Receptor-2 Signaling-Dependent Mouse Corneal Wound Healing.
PLoS ONE 8(5): e63674.
doi: 10.1371/journal.pone.0063674
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rat anti Mouse Ly6B.2 antibody, clone F7/4 (MCA771GA) used to identify neutrophils by immunofluorescence.
Image caption:
Inflammation observed in full thickness wounds is caused by neutrophils. (A–D) Confocal images of WT and Jam-Agt/gt eye sections. (C) Co-localization of MMP-9 and Ly-6B.2 depicted in WT and Jam-Agt/gt corneas of full thickness injury eyes. Scale bar 100 µm. Nuclear staining using Draq5 is shown in blue. CE: Corneal epithelium; CS: Corneal stroma.
From: Chatterjee S, Wang Y, Duncan MK, Naik UP (2013)
Junctional Adhesion Molecule-A Regulates Vascular Endothelial Growth Factor Receptor-2 Signaling-Dependent Mouse Corneal Wound Healing.
PLoS ONE 8(5): e63674.
doi: 10.1371/journal.pone.0063674
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rat anti Mouse Ly6B.2 antibody, clone F7/4 (MCA771GA) used to identify neutrophils by immunofluorescence.
Image caption:
Inflammation observed in full thickness wounds is caused by neutrophils. Confocal images of WT and Jam-Agt/gt eye sections. (D) Split images of MMP-9 and Ly-6B.2 staining of 12 day WT are also shown. Scale bar 10 µm. All inset magnifications are 10x of the highlighted region. Nuclear staining using Draq5 is shown in blue. CE: Corneal epithelium; CS: Corneal stroma.
From: Chatterjee S, Wang Y, Duncan MK, Naik UP (2013)
Junctional Adhesion Molecule-A Regulates Vascular Endothelial Growth Factor Receptor-2 Signaling-Dependent Mouse Corneal Wound Healing.
PLoS ONE 8(5): e63674.
doi: 10.1371/journal.pone.0063674
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 monoclonal antibody, clone 7/4 (MCA771G) used for the identification of neutrophils in mouse colon by immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
CHL1 deficiency induced inflammatory cell infiltration (A,B) Representative immunohistochemical images (A) and analysis (B) of Ly6B.2-stained colon tissue from DSS-exposed CHL1+/+ and CHL1−/− mice (*p <0.05, **p <0.01, n = 5/group).
From: Han Y, Wang X, Cheng X, Zhao M, Zhao T, Guo L, Liu D, Wu K, Fan M, Shi M, Zhu L.
Close Homolog of L1 Deficiency Exacerbated Intestinal Epithelial Barrier Function in Mouse Model of Dextran Sulfate Sodium-Induced Colitis.
Front Physiol. 2020 Nov 6;11:584508.
doi: 10.3389/fphys.2020.584508.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 antibody, clone 7/4 (MCA771G) used to detect Ly-6B.2 expressing cells in murine colon by immunofluorescence.
Image caption:
Juvenile SDS increased the number of 5-HT-positive cells. Representative images of immunohistochemistry for CD4-, CD68-, 5-HT-, Ly6B.2-, in the colonic mucosa of control and stress mice. The number of CD4-, CD68-, 5-HT-, Ly6B.2-positive cells per 106 μm2 were counted. Data are presented as the mean ± SEM (n = 7, 8 mice per group). *p< 0.05 compared with the control group. Scale bars, 50 μm.
From: Matsumoto K, Takata K, Yamada D, Usuda H, Wada K, Tada M, Mishima Y, Ishihara S, Horie S, Saitoh A, Kato S.
Juvenile social defeat stress exposure favors in later onset of irritable bowel syndrome-like symptoms in male mice.
Sci Rep. 2021 Aug 11;11(1):16276.
doi: 10.1038/s41598-021-95916-5.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rat anti mouse Ly6B.2 antibody, clone F7/4 (MCA771G) used to label neutrophils in mouse kidney by immunofluorescence.
Image caption:
Effect of Trpc6 deficiency on renal neutrophils infiltration and inflammatory markers in IRI–induced AKI.
(A) Representative outer medullar images of IRI-injured kidneys. Kidneys were stained with Ly6B.2 (magnification: 400 ×). Scale bars are 100μm. (B) The semi-quantification renal neutrophils infiltration (Sham WT and Trpc6−/− n = 3 each, IRI WT n = 6, and Trpc6−/− n = 6, respectively).
From: Zheng Z, Tsvetkov D, Bartolomaeus TUP, Erdogan C, Krügel U, Schleifenbaum J, Schaefer M, Nürnberg B, Chai X, Ludwig FA, N'diaye G, Köhler MB, Wu K, Gollasch M, Markó L.
Role of TRPC6 in kidney damage after acute ischemic kidney injury.
Sci Rep. 2022 Feb 22;12(1):3038.
doi: 10.1038/s41598-022-06703-9.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Rat anti Mouse Ly-6B.2 antinbody, clone F7/4 (MCA771G) used to label neutrophils in mouse aortic tissue by immunohistochemistry on formalin fixed, paraffin embedded samples.
Image caption:
Panx1 deletion on endothelial cells mitigates leukocyte trafficking in AAA.
a Comparative histology and immunohistochemistry performed on day 14 indicates a marked decrease in neutrophil (PMN), and macrophage (Mac-2) immunostaining in aortic tissue of elastase-treated EC-Panx1−/− mice compared with elastase-treated WT and SMC-Panx1−/− mice. n = 5/group. Arrows indicate areas of immunostaining. Scale bar is 200 μm.
From: Filiberto AC, Spinosa MD, Elder CT, Su G, Leroy V, Ladd Z, Lu G, Mehaffey JH, Salmon MD, Hawkins RB, Ravichandran KS, Isakson BE, Upchurch GR Jr, Sharma AK.
Endothelial pannexin-1 channels modulate macrophage and smooth muscle cell activation in abdominal aortic aneurysm formation.
Nat Commun. 2022 Mar 21;13(1):1521.
doi: 10.1038/s41467-022-29233-4.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Filter by Application:
IF C P F ResetRat anti Mouse Ly-6B.2 Alloantigen
- Product Type
- Monoclonal Antibody
- Clone
- 7/4
- Isotype
- IgG2a
- Specificity
- Ly-6B.2 Alloantigen
Quick Links:
Antibody Quality and 
Unrivaled 
| Rat anti Mouse Ly-6B.2 monoclonal antibody, clone 7/4 recognizes the Ly-6B.2 antigen. Ly-6B.2 is a ~25-30 kDa GPI-anchored, heavily glycosylated protein expressed on neutrophils, inflammatory monocytes and some activated macrophages (Rosas et al. 2010). High levels of expression are seen in bone marrow, spleen, lung and lymph nodes. N-glycanase treatment of thioglycollate elicited peritoneal neutrophil lysates lowers the apparent molecular weight of Ly-6B.2 to ~15 kDa (Rosas et al.2010). In common with other Ly-6 antigens Ly-6B.2 demonstrates a polymorphic expression on inbred mouse strains (Kimura et al. 1984). Rat anti mouse Ly-6B.2, clone 7/4 recognizes the Ly-6B.2 antigen in 129J; AKR; C57BL/6; C57BL/10; C58; DBA/2; NZB; NZW; SJL; MFI; Swiss (PO) Strains whilst A2G; A/Sn; ASW; BALB/c; C3H/HEH: CBA.T6T6 are negative or demonstrate very weak reactivity (Hirsch and Gordon 1982). Rat anti mouse Ly-6B.2 has been successfully used for the immunomagnetic depletion of neutrophils during the enrichment of primitive hematopoietic cells from bone marrow (Bertoncello et al. 1991) and the depletion of myeloid cells in vivo (Rosas et al. 2010). |
- Target Species
- Mouse
- Product Form
- Purified IgG - liquid
- Preparation
- MCA771G, MCA771GA: Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
- MCA771GT: Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant.
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
0.09% Sodium Azide - Carrier Free
- Yes
- Immunogen
- Cultured bone marrow cells
- Approx. Protein Concentrations
- Pack Size: 0.25 mg, 25 µgIgG concentration 1.0 mg/mlPack Size: 0.1 mgIgG concentration 1 mg/ml
- Fusion Partners
- Spleen cells from AO rats were fused with cells from the Y3 Ag1.2.3 rat myeloma cell line.
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
Store at +4oC. DO NOT FREEZE.
This product should be stored undiluted. Should this product contain a precipitate we recommend microcentrifugation before use.
This product should be stored undiluted. Should this product contain a precipitate we recommend microcentrifugation before use.
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| Flow Cytometry |  |
1/50 | 1/100 |
| Immunofluorescence | |
||
| Immunohistology - Frozen | |
||
| Immunohistology - Paraffin | |
||
| Western Blotting | |
Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use their own system using appropriate negative/positive controls.
- Flow Cytometry
- Use 10ul of the suggested working dilution to label 106 cells in 100ul.
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Rat IgG2a Negative Control | MCA1212 | E F | 1 ml |
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| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
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| Antigen Retrieval Buffer, pH8.0 | BUF025A | P | 500 ml | Log in | |||
| List Price | Your Price | ||||||
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| Description | Antigen Retrieval Buffer, pH8.0 | ||||||
Source Reference
-
Hirsch, S. & Gordon, S. (1983) Polymorphic expression of a neutrophil differentiation antigen revealed by monoclonal antibody 7/4.
Immunogenetics. 18 (3): 229-39.
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Hobbs, J.A. et al. (2003) Myeloid cell function in MRP-14 (S100A9) null mice.
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- RRID
- AB_322950
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