mAbs Against HUMAN Protein
CD8A
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | rat anti human |
| CLONE NAME: | NOR132 |
| PROTEIN: | Human full length CD8A protein |
| PROTEIN WEB: | http://www.ncbi.nlm.nih.gov/omim/186910 |
| ANTIGEN USED: | Full lenght HIS-CD8 |
| FUSION PARTNER: | myeloma p3-NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG2a |
| SPECIES REACTIVITY: | Human |
| PREPARATION AND STORAGE: | Aliquot and store at 4ºC. Do not freeze |
| APP RECOMMENDED: | IHQ-frozen, IHQ-paraffin, IF, WB, IP |
| APP NO TESTED: | Flow cytometry |
Description
CD8 (cluster of differentiation 8) is a transmembrane glycoprotein that serves as a co-receptor for the T cell receptor (TCR). Like
the TCR, CD8 binds to a major histocompatibility complex (MHC) molecule, but is specific for the class I MHC protein. CD8
antigen is strongly expressed on human cytotoxic T cells and thymocytes, and is also expressed on a subset of NK cells.
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | NOR132 | neat | supernatan | none | goar anti rat HRP DAKO1:100 | tonsil / | membrane | ||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | NOR132 | 1:20 | supernatan | 30min ER1 | Novolink kit | tonsil / pancreas | membrane | Available | |
| Immunofluorescence | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | NOR132 | neat | supernatan | Tris-EDTA | Goat anti mouse IgGra Alexa | tonsil / | membrane | Available | |
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- IHC of CD8 (NOR132H) and CD8 (C8/144B) in transfected cells.
- Antibody CD8 (NOR132H) was characterized by immunohistochemistry on transfected Hek-pcDNA3-V5-CD8 cells. This antibody specificity has been compared with anti-CD8 (clone C8/144B)
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- Expression of CD8 (NOR132) and CD8 (C8/144B) in frozen tonsil
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- Expression of CD8 (NOR132) and CD8 (C8/144B) on paraffin tonsil, spleen, testicle and pancreas
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- Expression of CD8 (NOR132) and CD8 (C8/144B) in paraffin tonsil using immunofluorescence technique
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | NOR132 | NEAT | supernatan | Tonsil / Jurkat | 30kDa | 30kDa | Human | Available |
Inconclusive Result obtained is not completely satisfactory. More testing are needed before using these reagents in this application | C8/144B | NEAT | supernatan | Tonsil / Jurkat | 30kDa | 30kDa | Human | Available |
| Immunoprecipitation | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | NOR132 | NEAT | supernatan | Tonsil / Jurkat | 30kDa | 30kDa | Human | Available |
Inconclusive Result obtained is not completely satisfactory. More testing are needed before using these reagents in this application | C8/144B | NEAT | supernatan | Tonsil / Jurkat | 30kDa | 30kDa | Human | Available |
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- Expression of CD8 (NOR132) and CD8 (C8/144B) by WB
- Lanes
Lane 1 Hek-CD8 (30цg) (+)
Lane 2 Hek-Bcl2 (30цg) (-)
Lane 3 Tonsil (100цg) (+)
Lane 4 Human Thymus (100цg) (+)
Lane 5 Human spleen (100цg) (+)
Lane 6 Mouse spleen (100цg) (-)
Lane 7 Peripheric blood (100цg) (+)
Lane 8 JURKAT (100цg) (-)
Lane 9 TOLEDO (100цg) (-)
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- IP of CD8 (NOR132) and CD8 (C8/144B) mAbs
- A. Immunoprecipitation of protein extracts from HEK-CD8, HEK-Bcl2, Tonsil and Jurkat with commercial CD8 mAb (clone C8/144B) (used as neat supernatant) followed by western blotting with CD8 NOR132H.
B. Immunoprecipitation of protein extracts from HEK-CD8, HEK-Bcl2, Tonsil and Jurkat with CD8 NOR132H (used as neat supernatant) followed by western blotting with CD8 NOR132H
C. Immunoprecipitation of protein extracts from HEK-CD8, HEK-Bcl2, tonsil and Jurkat with commercial CD8 mAb (clone C8/144B) (used as neat supernatant) followed by western blotting with CD8 C8/144B.
D. Immunoprecipitation of protein extracts from HEK-CD8, HEK-Bcl2, Tonsil and Jurkat with CD8 NOR132H (used as neat supernatant) followed by western blotting with CD8 C8/144B.