CD335 antibody | AKS1
Mouse anti bovine CD335 antibody, clone AKS1 (MCA2365GA) used for the isolation by immunomagnetic separation and evaluation of CD335 expression by flow cytometry of NKp46 expression on bovine peripheral blood mononuclear cells.
Image caption:
CD2 distribution on bovine NK cells in peripheral blood and during cell culture. (A) CD2 expression by PBMCs from healthy cattle, and in NKp46-selected cells cultured for the indicated time in the presence of 100U/ml rbIL-2. Results were obtained by two-colour flow cytometry, gating for viable lymphocytes. Data shown are from one animal representative of 19. (B) Monitoring of cell divisions of CD2- and CD2+ NK cell populations during three and six days of rbIL-2 (100U/ml) stimulation. NKp46-selected cells were labelled with CFSE the day after isolation and CFSE intensity (filled histograms) measured in flow cytometry on the indicated days, gating for CD2- or CD2+ viable cells. Solid line open histograms show non-dividing NK cells, cultured in the presence of 1U/ml rbIL-2, while broken line open histograms show unlabeled stimulated NK cells. Data shown are from one animal representative of five. (C) Stability of CD2 expression in NK cell subsets cultured separately. NK cells separated with immunomagnetic beads for absence or presence of CD2 were cultured separately for eleven days in the presence of IL-2, monitoring CD2 expression at three different time points (filled histograms). Day 1 corresponds to the day of separation, 24 h after primary NK cell isolation. Open histograms show secondary mAb controls. Data shown are from one animal representative of eight.
From: Boysen P, Olsen I, Berg I, Kulberg S, Johansen GM, Storset AK.
Bovine CD2-/NKp46+ cells are fully functional natural killer cells with a high activation status. BMC Immunol. 2006 Apr 27;7:10.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Bovine CD335 antibody, clone AKS1 (MCA2365EL) used as a positive antibody control in blocking experiments looking at lytic and γ interferon responses or bovine NK cells.
Image caption:
Effect of anti-CD2 mAbs on bovine NK cell functions. (A) IFN-γ response from NK cells caused by cross-linking of CD2. Plastic wells were pre-coated with two different mAbs against bovine CD2, along with NKp46-mAb as positive control and CD8-mAb or medium as negative controls. rbIL-2 activated NK cells were added and incubated for 24 h in the presence or absence of rbIL-12 (0.5 U/ml), and IFN-γ content in the supernatants analyzed by an ELISA. Data shown are means ± SD of two to four individual animals. (B-C) IL-2 activated NK cell killing of the bovine target lines BL3.1 and MDBK in a 4 h 51Cr-release assay, with addition of the indicated mAbs for blocking. CD2mAb I = BAQ95A, CD2 mAb II = MUC2A, CD8 mAb = CACT80C, NKp46 mAb = AKS1. (D) As above, using murine Fc-receptor bearing P815 target cells to obtain a redirected lysis effect. Data shown in (B-D) are from one animal representative of three to five.
From: Boysen P, Olsen I, Berg I, Kulberg S, Johansen GM, Storset AK.
Bovine CD2-/NKp46+ cells are fully functional natural killer cells with a high activation status.
BMC Immunol. 2006 Apr 27;7:10.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti bovine CD335 antibody, clone AKS1 (MCA2365GA) used for the evaluation of NKp46 expression on bovine peripheral blood mononuclear cells by flow cytometry
Image caption:
Oenothein B Primes bovine CD335+ cells to respond to IL-18. (A) Bovine PBMCs (105 cells/well) were depleted of CD335+ cells and treated with 20 μg/ml oenothein B or X-VIVO medium alone for 24 hrs. Cells were then washed and treated with 10 ng/ml rhu IL-18 or medium alone for 18 hrs. After incubation, IFNγ levels in the supernatant fluids were measured by ELISA. The data are expressed as mean +/− SEM of three independent experiments comparing depleted PBMCs to un-depleted controls tested concurrently. All samples were tested in duplicate or triplicate. Statistical significance was measured by Two-way ANOVA with Bonferroni post-test. *p<0.05, **p<0.01, ***p<0.001 (B) Bovine PBMCs (105 cells/well) from a new calf were treated with the indicated amounts of oenothein B or X-VIVO medium alone for 24 hrs. Cells were washed and treated with 10 ng/ml rhu IL-18 or X-VIVO medium alone for 6 hrs in the presence of brefeldin A. IFNγ production was measured by intracellular flow cytometry. The data are expressed as mean+/− SEM. All samples were tested in triplicate. Statistical significance was measured by Two-way ANOVA with Bonferroni post-test. *p<0.05, **p<0.01, ***p<0.001 (C) Representative examples of two-color flow cytometry plots comparing IFNγ staining on CD335+ cells.
From: Ramstead AG, Schepetkin IA, Quinn MT, Jutila MA (2012)
Oenothein B, a Cyclic Dimeric Ellagitannin Isolated from Epilobium angustifolium, Enhances IFNγ Production by Lymphocytes.
PLoS ONE 7(11): e50546.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti bovine CD335 antibody, clone AKS1 (MCA2365GA) used for the evaluation of NKp46 expression on bovine peripheral blood mononuclear cells by flow cytometry
Image caption:
Oenothein B induces IL-2Rα or CD69 on bovine and human NK cells. (A) Bovine PBMCs (105 cells/well) were treated with 20 μg/ml oenothein B in X-VIVO medium for 24 hrs, and IL-2Rα expression on NK cells was measured by multi-color flow cytometry. Representative examples of two-color flow cytometry plots comparing IL-2Rα staining on oenothein B-treated and untreated bovine NK cells (CD335+) from each animal are shown. (B) Human PBMCs (105 cells/well) were treated with 40 μg/ml oenothein B in cRPMI medium for 48 hrs. CD69 expression on NK cells was then measured by flow cytometry. Representative examples of two-color flow cytometry plots comparing CD69 staining on oenothein B-treated and untreated human NK cells from each donor are shown.
From: Ramstead AG, Schepetkin IA, Quinn MT, Jutila MA (2012)
Oenothein B, a Cyclic Dimeric Ellagitannin Isolated from Epilobium angustifolium, Enhances IFNγ Production by Lymphocytes.
PLoS ONE 7(11): e50546.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti CD355 antibody, clone AKS1 (MCA2365GA) used to evaluate NKp46 expression on bovine peripheral blood mononuclear cells by flow cytometry.
Image caption:
PBMC from a TB-reactor animal were stimulated with PPDB and flow cytometry analysis performed to enumerate IL-22 and IL-17A production in different lymphocyte populations. Numbers represent the percentage of single, live bovine lymphocytes. Data representative of at least 4 independent experiments.
From: Steinbach S, Vordermeier HM, Jones GJ.
CD4+ and γδ T Cells are the main Producers of IL-22 and IL-17A in Lymphocytes from Mycobacterium bovis-infected Cattle.
Sci Rep. 2016 Jul 18;6:29990.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Phycoerythrin conjugated Mouse anti Bovine CD335 antibody, clone AKS1 (MCA2365PE) used along with FITC conjugated Mouse anti Bovine CD2 antibody, clone CC42 (MCA833F) to evaluate NKp46 expression on bovine lymphocyte populations by flow cytometry.
Image caption:
CD2− natural killer (NK) cells are the predominant subset of NK cells present within skin‐draining afferent lymph (AL). Lymphocytes derived from peripheral blood (PB), AL and the lymph nodes (LNs) of seven calves were labelled with monoclonal antibodies to NKp46 and CD2 and analysed by flow cytometry. FACS plots from one representative animal illustrate the expression of NKp46 and CD2 by PB‐, AL‐ and LN‐derived lymphocytes.
From: Hamilton CA, Mahan S, Bell CR, Villarreal-Ramos B, Charleston B, Entrican G, Hope JC.
Frequency and phenotype of natural killer cells and natural killer cell subsets in bovine lymphoid compartments and blood.
Immunology. 2017 May;151(1):89-97.
doi: 10.1111/imm.12708.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Phycoerythrin conjugated Mouse anti Bovine CD335 antibody, clone AKS1 (MCA2365PE) used along with FITC conjugated Mouse anti Bovine CD62L antibody, clone CC32 (MCA1649F) to evaluate NKp46 expression on bovine lymphocyte populations by flow cytometry.
Image caption:
CD62L expression by afferent‐lymph (AL) ‐derived CD2− natural killer (NK) cells may permit homing to lymph nodes (LNs). Lymphocytes from peripheral blood (PB), AL and LNs from seven calves were labelled with monoclonal antibodies for NKp46 and CD62L and analysed by flow cytometry. FACS plots from one representative animal illustrate the expression of NKp46 and CD62L by PB‐, AL‐ and LN‐derived lymphocytes (a). Gates were set based on Fluorescence Minus One controls.
From: Hamilton CA, Mahan S, Bell CR, Villarreal-Ramos B, Charleston B, Entrican G, Hope JC.
Frequency and phenotype of natural killer cells and natural killer cell subsets in bovine lymphoid compartments and blood.
Immunology. 2017 May;151(1):89-97.
doi: 10.1111/imm.12708.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Alexa Fluor 488 conjugated Mouse anti Bovine CD335 antibody, clone AKS1 (MCA1655F) used to assess CD335 (NKp46) expression on bovine cells by flow cytometry.
Image caption:
Representative dot plots. Gate strategy: (1) R1, lymphocyte gate used for WC1+ cells and (2) R2, gate used for NKp46+ cells. Dot plots from representative samples of WC1+IFN-γ+ in line A and NKp46+ IFN-γ+ in line B for non-infected and infected co-cultures (double positive events are represented in the upper right quadrant of each dot plot). Representative dot plots of NKp46+IFN-γ+ cells incubated with 5mg and 10mg lyophilized bacterial culture medium supernatant (CS) from Mb04-303 are depicted in line C.
From: Blanco FC, Gravisaco MJ, Bigi MM, García EA, Marquez C, McNeil M, Jackson M, Bigi F.
Identifying Bacterial and Host Factors Involved in the Interaction of Mycobacterium bovis with the Bovine Innate Immune Cells.
Front Immunol. 2021 Jul 15;12:674643.
doi: 10.3389/fimmu.2021.674643.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Bovine CD335
- Product Type
- Monoclonal Antibody
- Clone
- AKS1
- Isotype
- IgG1
- Specificity
- CD335
Antibody Quality and Performance Guarantee
| Mouse anti Bovine CD335 antibody, clone AKS1 recognizes bovine CD335, also known as NKp46 and Natural cytotoxicity triggering receptor 1. CD335 is a type I transmembrane protein, with two extracellular C2-type immunoglobulin-like domains, which functions as an activating receptor. CD335 is expressed by human natural killer cells (Sivori et al. 1997). The bovine homologue is expressed on bovine NK cells (Storset et al. 2004) and no expression of CD335 has been detected on B cells, T cells, monocytes or granulocytes. Clone AKS1 is reported to activate lysis of FcgammaR-expressing cell line P815, by IL-2 activated NKp46+ cells (Storset et al. 2004). |
- Target Species
- Bovine
- Species Cross-Reactivity
-
Target Species Cross Reactivity American Bison - N.B. Antibody reactivity and working conditions may vary between species.
- Product Form
- Purified IgG - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.09% sodium azide (NaN3)
- Carrier Free
- Yes
- Immunogen
- Fusion protein consisting of the extracellular region of bovine CD335.
- Approx. Protein Concentrations
- IgG concentration 1.0 mg/ml
- Fusion Partners
- Spleen cells from immunised Balb/c mice were fused with cells of the NS-0 myeloma cell line.
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
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 | 1/200 |
| Immunofluorescence | |
||
| Western Blotting | |
Where this product 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 product for use in their own system using appropriate negative/positive controls.
- Flow Cytometry
- Use 10μl of the suggested working dilution to label 106 cells in 100μl
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Mouse IgG1 Negative Control | MCA928 | F | 100 Tests |
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| Description | Mouse IgG1 Negative Control | ||||||
References for CD335 antibody
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Storset, A.K. et al. (2004) NKp46 defines a subset of bovine leukocytes with natural killer cell characteristics.
Eur J Immunol. 34 (3): 669-76. -
Kulberg, S. et al. (2004) Reference values for relative numbers of natural killer cells in cattle blood.
Dev Comp Immunol. 28 (9): 941-8. -
Van Rhijn, I. et al. (2007) Massive, sustained γ/δ T cell migration from the bovine skin in vivo.
J Leukoc Biol. 81: 968-73. -
Toka, F.N. et al. (2011) Rapid and Transient Activation of γ/δ T Cells to IFN-γ Production, NK Cell-Like Killing, and Antigen Processing during Acute Virus Infection.
J Immunol. 186: 4853-61. -
Bastos, R.G. et al. (2008) Bovine NK cells acquire cytotoxic activity and produce IFN-γ after stimulation by Mycobacterium bovis BCG- or Babesia bovis-exposed splenic dendritic cells.
Vet Immunol Immunopathol. 124: 302-12. -
Boysen P et al. (2006) Bovine CD2-/NKp46+ cells are fully functional natural killer cells with a high activation status.
BMC Immunol. 7: 10. -
Elhmouzi-Younes, J. et al. (2009) Bovine neonate natural killer cells are fully functional and highly responsive to interleukin-15 and to NKp46 receptor stimulation.
Vet Res. 40: 54. -
Elhmouzi-Younes, J. et al. (2010) Ovine CD16+/CD14- blood lymphocytes present all the major characteristics of natural killer cells.
Vet Res. 41:4.
View The Latest Product References
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Hoek, A. et al. (2009) Subpopulations of bovine WC1(+) γ/δ T cells rather than CD4(+)CD25(high) Foxp3(+) T cells act as immune regulatory cells ex vivo.
Vet Res. 40:6. -
Zuerner, R.L. et al. (2011) A Leptospira borgpetersenii Serovar Hardjo Vaccine Induces a Th1 Response, Activates NK Cells, and Reduces Renal Colonization.
Clin Vaccine Immunol. 18: 684-91. -
Nelson, D.D. et al. (2010) CD8(+)/perforin(+)/WC1(- ) γ/δT cells, not CD8(+) α/β T cells, infiltrate vasculitis lesions of American bison (Bison bison) with experimental sheep-associated malignant catarrhal fever.
Vet Immunol Immunopathol. 136: 284-91. -
Banos, G. et al. (2013) Identification of immune traits correlated with dairy cow health, reproduction and productivity.
PLoS One. 8: e65766. -
Ramstead, A.G. et al. (2015) Aging influences the response of T cells to stimulation by the ellagitannin, oenothein B.
Int Immunopharmacol. 26 (2): 367-77. -
Boysen, P. et al. (2008) Natural killer cells in lymph nodes of healthy calves express CD16 and show both cytotoxic and cytokine-producing properties.
Dev Comp Immunol. 32 (7): 773-83. -
González-Cano, P. et al. (2014) Two functionally distinct myeloid dendritic cell subpopulations are present in bovine blood.
Dev Comp Immunol. 44 (2): 378-88. -
Di Palma, S. et al. (2012) Comparative spatiotemporal analysis of the intrathecal immune response in natural listeric rhombencephalitis of cattle and small ruminants.
Comp Immunol Microbiol Infect Dis. 35 (5): 429-41. -
Pirson, C. et al. (2015) Highly purified mycobacterial phosphatidylinositol mannosides drive cell-mediated responses and activate NKT cells in cattle.
Clin Vaccine Immunol. 22 (2): 178-84. -
Schneider, D.A. et al. (2011) Dynamics of bovine spleen cell populations during the acute response to Babesia bovis infection: an immunohistological study.
Parasite Immunol. 33 (1): 34-44. -
Van Meulder F et al. (2015) Analysis of the protective immune response following intramuscular vaccination of calves against the intestinal parasite Cooperia oncophora.
Int J Parasitol. 45 (9-10): 637-46. -
Patch, J.R. et al. (2014) Infection with foot-and-mouth disease virus (FMDV) induces a natural killer (NK) cell response in cattle that is lacking following vaccination.
Comp Immunol Microbiol Infect Dis. 37 (4): 249-57. -
Hedges, J.F. et al. (2015) Amphotericin B stimulates γδ T and NK cells, and enhances protection from Salmonella infection.
Innate Immun. 21 (6): 598-608. -
Allan, A.J. et al. (2015) Cattle NK Cell Heterogeneity and the Influence of MHC Class I.
J Immunol. 195 (5): 2199-206. -
Maślanka, T. et al. (2016) Prostaglandin E2 exerts the proapoptotic and antiproliferative effects on bovine NK cells
Res Vet Sci. 107: 80-7. -
Hecker YP et al. (2015) Cell mediated immune responses in the placenta following challenge of vaccinated pregnant heifers with Neospora caninum.
Vet Parasitol. 214 (3-4): 247-54. -
Ohira K et al. (2016) Bovine leukemia virus reduces anti-viral cytokine activities and NK cytotoxicity by inducing TGF-β secretion from regulatory T cells.
Immun Inflamm Dis. 4 (1): 52-63. -
Johnson, W.C. et al. (2008) Bovine WC1(-) γδT cells incubated with IL-15 express the natural cytotoxicity receptor CD335 (NKp46) and produce IFN-γ in response to exogenous IL-12 and IL-18.
Dev Comp Immunol. 32 (8): 1002-10. -
Sipka, A. et al. (2016) Bovine natural killer cells are present in Escherichia coli infected mammary gland tissue and show antimicrobial activity in vitro.
Comp Immunol, Microbiol Infect Dis. 48: 54-60. -
Hamilton, C.A. et al. (2016) Interactions between natural killer cells and dendritic cells favour T helper1-type responses to BCG in calves.
Vet Res. 47 (1): 85. -
Krueger, L.A. et al. (2016) Gamma delta T cells are early responders to Mycobacterium avium ssp. paratuberculosis in colostrum-replete Holstein calves.
J Dairy Sci. 99 (11): 9040-50. -
Steinbach, S. et al. (2016) CD4+ and γδ T Cells are the main Producers of IL-22 and IL-17A in Lymphocytes from Mycobacterium bovis-infected Cattle.
Sci Rep. 6: 29990. -
Rutigliano, H.M. et al. (2016) Trophoblast Major Histocompatibility Complex Class I Expression Is Associated with Immune-Mediated Rejection of Bovine Fetuses Produced by Cloning.
Biol Reprod. 95 (2): 39. -
González-Hernández A et al. (2016) Host protective ASP-based vaccine against the parasitic nematode Ostertagia ostertagi triggers NK cell activation and mixed IgG1-IgG2 response.
Sci Rep. 6: 29496. -
Khosa, S. et al. (2020) Bovine Adenovirus-3 Tropism for Bovine Leukocyte Sub-Populations.
Viruses. 12 (12): 1431. -
Denholm, S.J. et al. (2017) Estimating genetic and phenotypic parameters of cellular immune-associated traits in dairy cows.
J Dairy Sci. 100 (4): 2850-2862. -
Hamilton, C.A. et al. (2017) Frequency and phenotype of natural killer cells and natural killer cell subsets in bovine lymphoid compartments and blood.
Immunology. 151 (1): 89-97. -
Bassi, P.B. et al. (2018) Parasitological and immunological evaluation of cattle experimentally infected with Trypanosoma vivax.
Exp Parasitol. 185: 98-106. -
de Araújo, F.F. et al. (2019) Distinct immune response profile during Rhipicephalus. (Boophilus.) microplus. infestations of guzerat dairy herd according to the maternal lineage ancestry (mitochondrial DNA).
Vet Parasitol. 273: 36-44. -
Nakajima, N. et al. (2019) Effects of direct exposure to cold weather under grazing in winter on the physiological, immunological, and behavioral conditions of Japanese Black beef cattle in central Japan.
Anim Sci J. 90 (8): 1033-1041. -
Okino, C.H. et al. (2020) A polymorphic CD4 epitope related to increased susceptibility to Babesia bovis. in Canchim calves.
Vet Immunol Immunopathol. 230: 110132. -
de Araújo, F.F. et al. (2019) Distinct immune response profile during Rhipicephalus (Boophilus) microplus. infestations of guzerat dairy herd according to the maternal lineage ancestry (mitochondrial DNA).
Vet Parasitol. 273: 36-44. -
Colombatti Olivieri, M.A. et al. (2021) Evaluation of a virulent strain of Mycobacterium avium subsp. paratuberculosis used as a heat-killed vaccine.
Vaccine. 39 (51): 7401-7412. -
Blanco, F.C. et al. (2021) Identifying Bacterial and Host Factors Involved in the Interaction of Mycobacterium bovis with the Bovine Innate Immune Cells.
Front Immunol. 12: 674643.
Further Reading
-
Sivori, S. et al. (1997) p46, a novel natural killer cell-specific surface molecule that mediates cell activation.
J Exp Med. 186 (7): 1129-36. -
Storset, A.K. et al. (2003) Natural killer cell receptors in cattle: a bovine killer cell immunoglobulin-like receptor multigene family contains members with divergent signaling motifs.
Eur J Immunol. 33 (4): 980-90.
- Synonyms
- NKp46
- UniProt
- Q863H2
- Entrez Gene
- NCR1
- GO Terms
- GO:0005886 plasma membrane
- GO:0016021 integral to membrane
- GO:0004872 receptor activity
MCA2365GA
153584
160063
1801
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