mAbs Against MOUSE Protein
CD8a
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | rat anti mouse |
| CLONE NAME: | OTO94A |
| PROTEIN: | mouse T-cell surface glycoprotein CD8 alpha chain |
| PROTEIN WEB: | http://www.uniprot.org/uniprot/P01731 |
| ANTIGEN USED: | Transfected cells with pCMV6-MYC-DDK-mCD8a |
| FUSION PARTNER: | NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG2c |
| SPECIES REACTIVITY: | mouse |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
Description
Integral membrane glycoprotein that plays an essential role in the immune response and serves multiple functions in responses against both external and internal offenses. In T-cells, functions primarily as a coreceptor for MHC class I molecule:peptide complex. The antigens presented by class I peptides are derived from cytosolic proteins while class II derived from extracellular proteins. Interacts simultaneously with the T-cell receptor (TCR) and the MHC class I proteins presented by antigen presenting cells (APCs). In turn, recruits the Src kinase LCK to the vicinity of the TCR-CD3 complex. LCK then initiates different intracellular signaling pathways by phosphorylating various substrates ultimately leading to lymphokine production, motility, adhesion and activation of cytotoxic T-lymphocytes (CTLs). This mechanism enables CTLs to recognize and eliminate infected cells and tumor cells. In NK-cells, the presence of CD8A homodimers at the cell surface provides a survival mechanism allowing conjugation and lysis of multiple target cells. CD8A homodimer molecules also promote the survival and differentiation of activated lymphocytes into memory CD8 T-cells (obtained from Uniprot).
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | OTO94A | 1:200 | supernatant | mouse spleen / | membrane | ||||
| Immunofluorescence | |||||||||
Enlarge image
- CD8a antibody (OTO94A) in HEK transfected cells.
- Validation of OTO94A monoclonal antibody in transfected cells. Membrane staining on frozen cytospin preparations of transfected HEK293T-MYC-mCD8a cells using antibody OTO94A. Anti-myc antibody was used as positive control.
Enlarge image
- OTO94A mAb staining in mouse lymphoid tissues.
| 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 | OTO94A | undiluted supernatant | mouse thymus / | 27kDa | 27kDa | Available | ||
| Immunoprecipitation | ||||||||
Enlarge image
- Western Blotting of anti-mCD8a (OTO94A) using different cell extracts.
- WB analysis of cell line samples and mouse thymus extract for mCD8a.
Lane 1 HEK-MYC-mCD8a transfected cells (10ug) (+)
Lane 2 HEK-DRB1 transfected cells (10ug) (-)
Lane 3 Mouse thymus extract (100ug) (+)
Lane 4 SUP-T1 human cell line (100ug) (-)
Lane 5 RBL-1-mCD8a transfected cells (10ug) (+)