Product nameAnti-CD8 alpha antibody [144B]
See all CD8 alpha primary antibodies
DescriptionMouse monoclonal [144B] to CD8 alpha
Tested applicationsSuitable for: IHC-R, WB, Flow Cyt, IHC-Pmore details
Species reactivityReacts with: Human
Synthetic peptide corresponding to Human CD8 alpha (C terminal).
Database link: P01732
- IHC-P: Human tonsil tissue. Flow Cyt: Human peripheral blood lymphocytes.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.05% Sodium azide
Constituents: Tissue culture supernatant, 1% BSA
PurityTissue culture supernatant
Light chain typekappa
Our Abpromise guarantee covers the use of ab17147 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-R||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration.|
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-P||1/25 - 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
FunctionIdentifies cytotoxic/suppressor T-cells that interact with MHC class I bearing targets. CD8 is thought to play a role in the process of T-cell mediated killing. CD8 alpha chains binds to class I MHC molecules alpha-3 domains.
Involvement in diseaseDefects in CD8A are a cause of familial CD8 deficiency (CD8 deficiency) [MIM:608957]. Familial CD8 deficiency is a novel autosomal recessive immunologic defect characterized by absence of CD8+ cells, leading to recurrent bacterial infections.
Sequence similaritiesContains 1 Ig-like V-type (immunoglobulin-like) domain.
modificationsAll of the five most carboxyl-terminal cysteines form inter-chain disulfide bonds in dimers and higher multimers, while the four N-terminal cysteines do not.
Cellular localizationSecreted and Cell membrane.
- Information by UniProt
- alpha polypeptide (p32) antibody
- CD8 antibody
- CD8 antigen alpha polypeptide antibody
Formalin fixed paraffin embedded human tonsil stained with CD8 using ABC and DAB chromogen.
Human peripheral blood lymphocytes stained with ab17147 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (ab17147, 1/100 dilution) for 30 min at 4°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy - peripheral blood lymphocytes.
ab17147 staining CD8 in human nasal polyp and tonsil tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with 10% formalin for 24 hours and a heat mediated antigen retrieval step was performed using EDTA pH 8.0. Samples were then blocked with 20% DMEM and 1% BSA in culture medium for 20 minutes at 21°C followed by incubation with the primary antibody at a 1/150 dilution, for 18 hours at 4°C. A biotinylated rabbit anti-mouse (Fab2) polyclonal was used as secondary antibody at a 1/400 dilution.An Avidin Biotin blocking step was used in addition to those already mentioned.
All lanes : Anti-CD8 alpha antibody [144B] (ab17147) at 1/500 dilution
Lane 1 : Peripheral T-lymphocyte lysate
Lane 2 : PC3 whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : AP-conjugated polyclonal goat anti-mouse IgG at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 29 kDa why is the actual band size different from the predicted?
Exposure time: 15 minutes
Blocked with 4% BSA for 2 hours at 20°C.
Incubated with the primary antibody for 30 minutes.
This product has been referenced in:
- Liu B et al. PD-1/PD-L1 expression in a series of intracranial germinoma and its association with Foxp3+ and CD8+ infiltrating lymphocytes. PLoS One 13:e0194594 (2018). Read more (PubMed: 29617441) »
- Nakamura M et al. Heterogeneity of programmed death-ligand expression in a case of Merkel cell carcinoma exhibiting complete regression after multiple metastases. Br J Dermatol N/A:N/A (2018). Read more (PubMed: 30431150) »