CD14 antibody | MIL2

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Anti Pig CD14 Antibody, clone MIL2 thumbnail image 1

Anti Pig CD14 Antibody, clone MIL2 thumbnail image 2

Anti Pig CD14 Antibody, clone MIL2 thumbnail image 3

Anti Pig CD14 Antibody, clone MIL2 thumbnail image 4

Anti Pig CD14 Antibody, clone MIL2 thumbnail image 5

Anti Pig CD14 Antibody, clone MIL2 gallery image 1 — Staining of pig peripheral blood granulocytes with Mouse anti Pig CD14 antibody, clone MIL2 (**MCA1218GA**) followed by FITC conjugated Goat anti Mouse IgG antibody (STAR117F)

Anti Pig CD14 Antibody, clone MIL2 gallery image 2 — **Published customer image:**
Mouse anti Pig CD14 antibody, clone MIL2 (**MCA1218GA**) used for the evaluation of CD14 expression on porcine leukocytes by flow cytometry.
**Image caption:**
Overview of flow cytometry analyses. (A) Gating strategy for identification of monocytes and macrophages. The peripheral blood is shown. Designation of population shown within each dot-plot is indicated above the dot-plot. Leukocytes were identified as viable (a) non-doublet (b) cells with typical light scatter properties of leukocytes (c). Then, macrophages were gated simply as CD203a^hi leukocytes (d) and marked with blue color. Monocytes were gated as CD203a^low/- SWC8^-(e) CD172a^hi(f) leukocytes where the CD203a^low/- region was defined as the complementary region to the CD203a^hi region. Then, SLA-DR+ monocytes were marked with red color and SLA-DR- monocytes were marked with green color (g). SLA-DR- region was defined as the complementary region to the SLA-DR+ region. Gating order is shown in the scheme (h). (B) Representative CD163 vs. CD14 dot-plots of macrophages (blue) and monocyte subpopulations (green: SLA-DR–, red: SLA-DR+) in various body compartments of control and APP-infected pigs.

From: Ondrackova P, Leva L, Kucerova Z, Vicenova M, Mensikova M, Faldyna M. Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental *Actinobacillus pleuropneumoniae* infection and the role of chemokines.
Vet Res. 2013 Oct 17;44:98.

Anti Pig CD14 Antibody, clone MIL2 gallery image 3 — **Published customer image:**
Mouse anti Pig CD14, clone MIL2 used for the separation of mococytes from porcine PBMCs by immunomagnetic cell sorting.
**Image caption:**
Porcine MoDCs at different culture times (400×).CD14⁺ monocytes were purified from porcine PBMCs and were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 25 ng/mL recombinant porcine IL-4, and 10 ng/ml recombinant porcine GM-CSF. The culture medium was replenished every 3 days. (A) CD14⁺ monocytes after 2 days of culture. (B) CD14⁺ monocytes 3 days of culture. (C) CD14⁺ monocytes after 4 days of culture. (D) CD14⁺ monocytes after 5 days of culture.

From: Citation: Liu J, Tian Z-Y, Xiao Y-C, Wang X-L, Jin M-L, Shi D-S (2016) The Role of Porcine Monocyte Derived Dendritic Cells (MoDC) in the Inflammation Storm Caused by *Streptococcus suis* Serotype 2 Infection. PLoS ONE 11(3): e0151256.

Anti Pig CD14 Antibody, clone MIL2 gallery image 4 — Figure A. FITC conjugated mouse anti porcine CD14 (**MCA1218F**) and A647 conjugated mouse IgG1 isotype control (MCA928A647). Figure B. FITC conjugated mouse anti porcine CD14 (**MCA1218F**) and A647 conjugated mouse anti porcine CD45 (MCA1222A647). All experiments performed on red cell lysed porcine blood gated on mononuclear cells.

Anti Pig CD14 Antibody, clone MIL2 gallery image 5 — Figure A. A647 conjugated mouse anti porcine CD45 (MCA1222A647) and FITC conjugated mouse IgG2b isotype control (MCA691F). Figure B. A647 conjugated mouse anti porcine CD45 (MCA1222A647) and FITC conjugated mouse anti porcine CD14 (**MCA1218F**). All experiments performed on red cell lysed porcine blood gated on mononuclear cells.

Mouse anti Pig CD14:FITC

Mouse anti Pig CD14

Product Type: Monoclonal Antibody
Clone: MIL2
Isotype: IgG2b

Product Code	Applications	Datasheet	MSDS	Pack Size	List Price	Quantity
MCA1218F	F			0.1 mg
MCA1218GA	C F IF			0.1 mg

Mouse anti Pig CD14, clone MIL2 recognizes porcine CD14. Clone MIL2 was clustered as porcine CD14 at the Third International Workshop on Swine Leukocyte Differentiation Antigens (Haverson et al. 2001) . Mouse anti Pig CD14, clone MIL2 immunoprecipitates a protein of ~50 kDa consistent with the expected apparent molecular weight of porcine CD14, and demonstrates the expected CD14 profile by dual labelling and competition studies. Further, pre-incubation of peripheral blood monocytes with MIL2 inhibits the binding of FITC labelled LPS, consistent with masking the CD14 LPS binding site (Thacker et al. 2001) .

Mouse anti pig CD14, clone MIL2 demonstrates staining of both monocytes and neutrophils in peripheral blood by flow cytometry with a similar expression pattern to the anti human CD14 clone TüK4, lymphocytes and eosinophils are negative for MIL2 staining (Zelnickova et al. 2007). Cloning and characterization of porcine CD14 indicates a high degree of both functional and structural conservation when compared to CD14 from other mammalian species, the gene maps to chromosome 2 and is expressed on a wide range of tissues in a manner consistent with expression on myeloid cells. (Petersen et al. 2007, Sanz et al. 2007).

Product Details

Target Species

Pig

Species Cross-Reactivity

Target Species	Cross Reactivity
Human

N.B. Antibody reactivity and working conditions may vary between species.

Product Form

Purified IgG conjugated to Fluorescein Isothiocyanate Isomer 1 (FITC) - liquid

Product Form

Purified IgG - liquid

Preparation

Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant

Preparation

Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant

Buffer Solution

Phosphate buffered saline

Buffer Solution

Phosphate buffered saline

Preservative Stabilisers

0.09%	Sodium Azide (NaN₃)
1%	Bovine Serum Albumin

Preservative Stabilisers

0.09% Sodium Azide (NaN₃)

Carrier Free

Yes

Immunogen

Porcine peripheral blood lymphocytes.

Approx. Protein Concentrations

IgG concentration 0.1mg/ml.

Approx. Protein Concentrations

IgG concentration 1.0 mg/ml

Fusion Partners

Spleen cells from immunized Balb/c mice were fused with cells from the P2-X63-Ag.653 mouse myeloma.

Storage Information

Storage: Store at +4^oC or at -20^oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted.
This product is photosensitive and should be protected from light.
Avoid repeated freezing and thawing as this may denature the antibody.
Should this product contain a precipitate we recommend microcentrifugation before use.
Storage: Store at +4^oC or at -20^oC if preferred.

This product should be stored undiluted.

Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Guarantee: 18 months from date of despatch.
Guarantee: 18 months from date of despatch.

More Information

UniProt: A2SW51 Related reagents
Regulatory: For research purposes only

Applications of CD14 antibody

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	Min Dilution	Max Dilution
Flow Cytometry
Flow Cytometry	1/25	1/200
Immunofluorescence
Immunohistology - Frozen

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 in their own system using appropriate negative/positive controls.

Flow Cytometry: Use 10ul of the suggested working dilution to label 10⁶ cells in 100ul.
Flow Cytometry: Use 10ul of the suggested working dilution to label 10⁶ cells in 100ul.

Secondary Antibodies Available

Description	Product Code	Pack Size	Applications
Human anti Mouse IgG2b:FITC	HCA038F	0.1 mg	F
Goat anti Mouse IgG (H/L):Alk. Phos. (Multi Species Adsorbed)	STAR117A	0.5 mg	E WB
Goat anti Mouse IgG (H/L):DyLight®488 (Multi Species Adsorbed)	STAR117D488GA	0.1 mg	F IF
Goat anti Mouse IgG (H/L):DyLight®680 (Multi Species Adsorbed)	STAR117D680GA	0.1 mg	F WB
Goat anti Mouse IgG (H/L):DyLight®800 (Multi Species Adsorbed)	STAR117D800GA	0.1 mg	F IF WB
Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed)	STAR117F	0.5 mg	F
Goat anti Mouse IgG (H/L):HRP (Multi Species Adsorbed)	STAR117P	0.5 mg	E WB
Goat anti Mouse IgG (Fc):FITC	STAR120F	1 mg	C F
Goat anti Mouse IgG (Fc):HRP	STAR120P	1 mg	E WB
Rabbit F(ab')2 anti Mouse IgG:RPE	STAR12A	1 ml	F
Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed)	STAR13B	1 mg	C E P RE WB
Goat anti Mouse IgG:FITC (Rat Adsorbed)	STAR70	0.5 mg	F
Goat anti Mouse IgG:RPE (Rat Adsorbed)	STAR76	1 ml	F
Goat anti Mouse IgG:HRP (Rat Adsorbed)	STAR77	0.5 mg	C E P
Goat anti Mouse IgG/A/M:Alk. Phos.	STAR87A	1 mg	C E WB
Goat anti Mouse IgG/A/M:HRP (Human Adsorbed)	STAR87P	1 mg	E
Rabbit F(ab')2 anti Mouse IgG:Dylight®800	STAR8D800GA	0.1 mg	F IF WB
Rabbit F(ab')2 anti Mouse IgG:FITC	STAR9B	1 mg	F

Negative Isotype Controls Available

Description	Product Code	Pack Size	Applications	List Price	Quantity
Mouse IgG2b Negative Control:FITC	MCA691F	100 Tests	F
Mouse IgG2b Negative Control	MCA691	100 Tests	F

Application Based External Images

Flow Cytometry

Functional Assays

Immunohistology - Frozen

Product Specific References

Source Reference

Haverson, K. et al. (1994) Characterization of monoclonal antibodies specific for monocytes, macrophages and granulocytes from porcine peripheral blood and mucosal tissues.
J Immunol Methods. 170 (2): 233-45.

References for CD14 antibody

Hauet, T. et al. (2000) Trimetazidine reduces renal dysfunction by limiting the cold ischemia/reperfusion injury in autotransplanted pig kidneys.
J Am Soc Nephrol. 11: 138-48.
Thacker, E. et al. (2001) Summary of workshop findings for porcine myelomonocytic markers.
Vet Immunol Immunopathol. 80 (1-2): 93-109.
Thorgersen, E.B. et al. (2010) CD14 inhibition efficiently attenuates early inflammatory and hemostatic responses in Escherichia coli sepsis in pigs.
FASEB J. 24: 712-22.
Goujon, J.M. et al. (2000) Influence of cold-storage conditions on renal function of autotransplanted large pig kidneys.
Kidney Int. 58: 838-50.
Li, Y. et al. (2014) Identification of apoptotic cells in the thymus of piglets infected with highly pathogenic porcine reproductive and respiratory syndrome virus.
Virus Res. 189: 29-33.
Summerfield, A. et al. (2003) Porcine peripheral blood dendritic cells and natural interferon-producing cells.
Immunology. 110: 440-9.
Vanderheijden, N. et al. (2003) Involvement of sialoadhesin in entry of porcine reproductive and respiratory syndrome virus into porcine alveolar macrophages.
J Virol. 77: 8207-15.
Barratt-Due, A. et al. (2011) Ornithodoros moubata Complement Inhibitor Is an Equally Effective C5 Inhibitor in Pigs and Humans.
J Immunol. 187: 4913-9.
Hauet, T. et al. (2002) Polyethylene glycol reduces the inflammatory injury due to cold ischemia/reperfusion in autotransplanted pig kidneys.
Kidney Int. 62: 654-67.
Kapetanovic, R. et al. (2012) Pig bone marrow-derived macrophages resemble human macrophages in their response to bacterial lipopolysaccharide.
J Immunol. 188: 3382-94.
Thorgersen, E.B. et al. (2009) Inhibition of complement and CD14 attenuates the Escherichia coli-induced inflammatory response in porcine whole blood.
Infect Immun. 77: 725-32.
Zelnickova, P. et al. (2007) Intracellular cytokine detection by flow cytometry in pigs: fixation, permeabilization and cell surface staining.
J Immunol Methods. 327: 18-29.
Facci, M.R. et al. (2011) Stability of expression of reference genes in porcine peripheral blood mononuclear and dendritic cells.
Vet Immunol Immunopathol. 141: 11-5.
Koutná, I. et al. (2012) Flow Cytometry Analysis of Intracellular Protein
In: Flow Cytometry - Recent Perspectives, Schmid, I. (Ed.), ISBN: 978-953-51-
Facci, M.R. et al. (2010) A comparison between isolated blood dendritic cells and monocyte-derived dendritic cells in pigs.
Immunology. 129: 396-405.
Schierack, P. et al. (2009) Effects of Bacillus cereus var. toyoi on immune parameters of pregnant sows.
Vet Immunol Immunopathol.127: 26-37.
Lundeland, B. et al. (2011) Severe gunshot injuries in a porcine model: impact on central markers of innate immunity.
Acta Anaesthesiol Scand. 55: 28-34.
Thorgersen, E.B. et al. (2008) Cyanobacterial LPS antagonist (CyP)-a novel and efficient inhibitor of Escherichia coli LPS-induced cytokine response in the pig.
Mol Immunol. 45: 3553-7.
Schierack, P. et al. (2007) Bacillus cereus var. toyoi enhanced systemic immune response in piglets.
Vet Immunol Immunopathol. 118: 1-11.
Ondrackova, P. et al. (2012) Interaction of porcine neutrophils with different strains of enterotoxigenic Escherichia coli.
Vet Microbiol. 160: 108-16.
Ondrackova, P. et al. (2013) Phenotypic characterisation of the monocyte subpopulations in healthy adult pigs and Salmonella-infected piglets by seven-colour flow cytometry.
Res Vet Sci. 94 (2): 240-5.
Vicenova, M. et al. (2014) Evaluation of in vitro and in vivo anti-inflammatory activity of biologically active phospholipids with anti-neoplastic potential in porcine model.
BMC Complement Altern Med. 14: 339.
Alvarez, B. et al. (2015) Phenotypic and functional heterogeneity of CD169⁺ and CD163⁺ macrophages from porcine lymph nodes and spleen.
Dev Comp Immunol. 44: 44-9.
Moffat, L. et al. (2014) Development and characterisation of monoclonal antibodies reactive with porcine CSF1R (CD115).
Dev Comp Immunol. 47 (1): 123-8.
Kyrova K et al. (2014) The response of porcine monocyte derived macrophages and dendritic cells to Salmonella typhimurium and lipopolysaccharide.
BMC Vet Res. 10: 244.
Nguyen, D.N. et al. (2016) Oral antibiotics increase blood neutrophil maturation and reduce bacteremia and necrotizing enterocolitis in the immediate postnatal period of preterm pigs.
Innate Immun. 22 (1): 51-62.
Egge, K.H. et al. (2015) Organ inflammation in porcine Escherichia coli sepsis is markedly attenuated by combined inhibition of C5 and CD14.
Immunobiology. 220 (8): 999-1005.
Liu J et al. (2016) The Role of Porcine Monocyte Derived Dendritic Cells (MoDC) in the Inflammation Storm Caused by Streptococcus suis Serotype 2 Infection.
PLoS One. 11 (3): e0151256.
Singleton, H. et al. (2016) Establishing Porcine Monocyte-Derived Macrophage and Dendritic Cell Systems for Studying the Interaction with PRRSV-1.
Front Microbiol. 7: 832.
Zemankova, N. et al. (2016) Bovine lactoferrin free of lipopolysaccharide can induce a proinflammatory response of macrophages.
BMC Vet Res. 12 (1): 251.
Auray, G. et al. (2016) Characterization and Transcriptomic Analysis of Porcine Blood Conventional and Plasmacytoid Dendritic Cells Reveals Striking Species-Specific Differences.
J Immunol. Nov 11. pii: 1600672. [Epub ahead of print]
Kavanová L et al. (2017) Concurrent infection with porcine reproductive and respiratory syndrome virus and Haemophilus parasuis in two types of porcine macrophages: apoptosis, production of ROS and formation of multinucleated giant cells.
Vet Res. 48 (1): 28.
Bacou, E. et al. (2017) β2-adrenoreceptor stimulation dampens the LPS-induced M1 polarization in pig macrophages.
Dev Comp Immunol. 76: 169-76.
Yang, G. et al. (2017) Characterizing porcine invariant natural killer T cells: A comparative study with NK cells and T cells.
Dev Comp Immunol. 76: 343-351.
Uitterdijk, A. et al. (2017) Time course of VCAM-1 expression in reperfused myocardial infarction in swine and its relation to retention of intracoronary administered bone marrow-derived mononuclear cells.
PLoS One. 12 (6): e0178779.
Sá:nchez, E.G. et al. (2017) Phenotyping and susceptibility of established porcine cells lines to African Swine Fever Virus infection and viral production.
Sci Rep. 7 (1): 10369.
Fernández-Caballero, T. et al. (2018) Phenotypic and functional characterization of porcine bone marrow monocyte subsets.
Dev Comp Immunol. 81: 95-104.
Sautter, C.A. et al. (2018) Phenotypic and functional modulations of porcine macrophages by interferons and interleukin-4.
Dev Comp Immunol. 84: 181-92.
López, E. et al. (2019) Identification of very early inflammatory markers in a porcine myocardial infarction model.
BMC Vet Res. 15 (1): 91.