CD335 antibody | EC1.1
Mouse anti Sheep CD335 antibody, clone EC1.1 (MCA5933GA) used to demonstrate specificity of the antibody by flow cytometry.
Image caption:
EC1.1 is specific for ovine NKp46. P815 cells transfected with an ovNKp46-FLAG construct are stained similarly by supernatant from EC1.1 (blue line) and a FLAG-specific antibody (red line) but not by an isotype control antibody (black line). Untransfected P815 cells are not stained by EC1.1 supernatant (solid grey).
From: Connelley T, Storset AK, Pemberton A, MacHugh N, Brown J, Lund H, Morrison IW.
NKp46 defines ovine cells that have characteristics corresponding to NK cells.
Vet Res. 2011 Feb 23;42(1):37.
doi: 10.1186/1297-9716-42-37.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Sheep CD335 antibody, clone EC1.1 (MCA5933GA) used to assess NKp46 expression on ovine cell populations by flow cytometry.
Image caption:
Analysis of the phenotype of ovine NKp46+ populations. (A)Two-colour flow cytometry analysis of ovine PBMC co-stained with EC1.1 and a selection of monoclonal antibodies against CD2, CD4, CD8, CD11b, CD16, WC1, IgM and MHCII. WC1 and IgM are surface molecules expressed by subsets of γδ T-cells and B-cells respectively. The results shown are representative of the results obtained from a minimum of five animals.
From: Connelley T, Storset AK, Pemberton A, MacHugh N, Brown J, Lund H, Morrison IW.
NKp46 defines ovine cells that have characteristics corresponding to NK cells.
Vet Res. 2011 Feb 23;42(1):37.
doi: 10.1186/1297-9716-42-37.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Sheep CD335 antibody, clone EC1.1 (MCA5933GA) used to assess NKp46 expression on ovine lymph node cell populations by flow cytometry.
Image caption:
Analysis of the phenotype of ovine NKp46+ populations. Two-colour flow cytometry analysis of mesenteric and pre-scapular lymph nodes and the spleen cell populations co-stained with EC1.1 and monoclonal antibodies against CD2 and CD8. The results shown are representative of the results obtained from a minimum of five animals.
From: Connelley T, Storset AK, Pemberton A, MacHugh N, Brown J, Lund H, Morrison IW.
NKp46 defines ovine cells that have characteristics corresponding to NK cells.
Vet Res. 2011 Feb 23;42(1):37.
doi: 10.1186/1297-9716-42-37.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Sheep CD335 antibody, clone EC1.1 (MCA5933GA) used to assess NKp46 expression in ovine lymph nodes and musosal tissues by immunofluorescence.
Image caption:
Distribution of ovine NKp46+ cells in lymph nodes and mucosal surfaces. Sections from ovine pre-scapular lymph node (A - cortex and B - medullary zone), abomasum (C), duodenum (D), ileum (E), colon (F), uterus (G) and lung (H). NKp46+ cells: green; B-cells (IgM+) red; CD4+ T-cells blue in panel A and B; Epithelial cells (E-cadherin+) blue in parts C-H. White arrows: individual NKp46+ cells. Red arrows: putative intra-epithelial NKp46+ cells. "Fo": follicle and "P": paracortex. "L": lymphocytic aggregate. Isotype matched controls demonstrated diffuse and dull background fluorescence in both the red and green channels (data not shown), however this could easily be distinguished by character and/or intensity from the bright cell-surface staining seen with the NKp46 and IgM-specific antibodies. Autofluorescent artifacts such as those demonstrated by the yellow arrows (e.g. in C and D) were not uncommon in the tissues examined. A scale for each of the figure parts is given. Images are taken from tissues that were representative of samples taken from a minimum of three animals as determined by preliminary single colour immuncytochemistry analysis.
From: Connelley T, Storset AK, Pemberton A, MacHugh N, Brown J, Lund H, Morrison IW.
NKp46 defines ovine cells that have characteristics corresponding to NK cells.
Vet Res. 2011 Feb 23;42(1):37.
doi: 10.1186/1297-9716-42-37.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
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- Product Type
- Monoclonal Antibody
- Clone
- EC1.1
- Isotype
- IgG1
- Specificity
- CD335
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Antibody Quality and 
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| Mouse anti Sheep CD335, clone EC1.1, recognizes the ovine homologue of human 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 function as an activating receptor. Research has indicated that, in mammals, the CD335 antigen is confined to NK cells (Westergard et. al. 2004) and that it is primarily expressed by a subset of natural killer (NK) cells which are CD3-ve, CD4-ve and CD2+ve with a small population also being CD8+ve (Storset et. al. 2004). No expression of CD335 has been detected on B cells, T cells, monocytes or granulocytes. NK cells are cytotoxic and form a key component of the innate immune system where they are involved in mediating immunity to tumours and pathogens and in particular are critical to the process where the host recognizes and rejects tumour and viral infected cells. Clone EC1.1 (originally named as Gr13.1) is the first antibody specifically raised to ovine CD335 and provides a reagent enabling researchers to identify ovine NKp46 positive NK cells and may facilitate an increased understanding of the ovine immune system and in particular its role in host pathogen defence (Connelley et. al. 2011). In addition to clone EC1.1, clones AKS1, which recognises CD335 (NKp46) in bovine and other ruminants, and VIV-KM1, which recognizes porcine CD335, are also available from Bio-Rad. |
- Target Species
- Sheep
- Species Cross-Reactivity
-
Target Species Cross Reactivity Bovine Goat - 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 A from tissue culture supernatant
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.09% Sodium Azide (NaN3)
- Carrier Free
- Yes
- Immunogen
- OvNKp46-mFcγ2b fusion protein
- Approx. Protein Concentrations
- IgG concentration 1.0 mg/ml
- Fusion Partners
- Spleen cells from immunised Balb/c mice were fused with cells of the X63 myeloma cell line
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
- Acknowledgements
- The Renaissance TSA Biotin system is a registered trademark of PerkinElmer, Waltham, USA
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 |  |
1/25 | 1/200 |
| Immunohistology - Frozen | |
||
| Immunohistology - Paraffin 1 | |
- 1 Mouse anti Sheep CD335 antibody, clone EC1.1 is suitable for use on paraffin sections when fixed with zinc salts Results may be enhanced by the use of a using TSA (Tyramide Signal Amplification) biotin signal enhancement system.
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 10ul of the suggested working dilution to label 106 cells in 100ul
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References for CD335 antibody
-
Connelley, T. et al. (2011) NKp46 defines ovine cells that have characteristics corresponding to NK cells.
Vet Res. 42: 37. -
Olsen, L. et al. (2013) Characterization of NCR1+ cells residing in lymphoid tissues in the gut of lambs indicates that the majority are NK cells
Vet Res. 44:109. -
Goh, S. et al. (2016) Identification of Theileria lestoquardi Antigens Recognized by CD8+ T Cells.
PLoS One. 11 (9): e0162571. -
Marzo, S. et al. (2021) Characterisation of dendritic cell frequency and phenotype in bovine afferent lymph reveals kinetic changes in costimulatory molecule expression
Vet Immunol Immunopathol. 19 Nov: 110363. -
Kolar, Q.K. et al. (2020) Anatomical distribution of respiratory tract leukocyte cell subsets in neonatal calves.
Vet Immunol Immunopathol. 227: 110090.
Further Reading
-
Elhmouzi-Younes, J. et al. (2010) Ovine CD16+/CD14- blood lymphocytes present all the major characteristics of natural killer cells.
Vet Res. 41:4 -
Storset, A. K. et. al. (2004) NKp46 defines a subset of bovine leukocytes with natural killer cell characteristics.
Eur J Immunol. 34:669-676 -
Westgaard, I.H. et al. (2004) Rat NKp46 activates natural killer cell cytotoxicity and is associated with FcepsilonRIgamma and CD3zeta.
J Leukoc Biol. 76 (6): 1200-6. -
Boysen, P. et al. (2011) Natural killer cells in free-living Mus musculus have a primed phenotype.
Mol Ecol. 20 (23): 5103-10.
- Synonyms
- Gr13.1
- NKp46
- UniProt
- E5LDC2
- Q863H2
- Entrez Gene
- NCR1
- GO Terms
- GO:0005886 plasma membrane
- GO:0016021 integral to membrane
- GO:0004872 receptor activity
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