Recombinant Anti-CD8 alpha antibody [EPR22483-288] (ab245118)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22483-288] to CD8 alpha
- Suitable for: IHC-P, Flow Cyt, IP, mIHC
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-CD8 alpha antibody [EPR22483-288]
See all CD8 alpha primary antibodies -
Description
Rabbit monoclonal [EPR22483-288] to CD8 alpha -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, Flow Cyt, IP, mIHCmore details
Unsuitable for: ICC/IF or WB -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- IHC-P: Human colon and tonsil tissue. Flow Cyt: Human peripheral blood mononuclear cells. IP: CD8 alpha IP in human thymus lysate.
-
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22483-288 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Immunohistochemistry reagents
-
Isotype control
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab245118 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
IHC-P | (1) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
Flow Cyt |
1/400.
|
|
IP |
1/30.
|
|
mIHC |
1/500.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) |
Notes |
---|
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt
1/400. |
IP
1/30. |
mIHC
1/500. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) |
Target
-
Function
Identifies 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 disease
Defects 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 similarities
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
Post-translational
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 localization
Secreted and Cell membrane. - Information by UniProt
-
Database links
- Entrez Gene: 925 Human
- Omim: 186910 Human
- SwissProt: P01732 Human
- Unigene: 85258 Human
-
Alternative names
- alpha polypeptide (p32) antibody
- CD_antigen=CD8a antibody
- CD8 antibody
see all
Images
-
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD8 alpha with ab245118 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on the T cell zone of human tonsil (PMID: 16504163) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
Immunohistochemical analysis of paraffin-embedded human colon tissue labeling CD8 alpha with ab245118 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on lymphocytes of human colon (PMID: 26310568) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human spleen tissue labeling CD8 alpha with ab245118 at 1/500 dilution, CD4 with ab238798 at 1/500, and CD19 with ab237772 at 1/5000 dilution.
Panel A: merged staining of anti-CD8 alpha (magenta; Opal™690), anti-CD4 (green; Opal™520) and anti-CD19 (red; Opal™570) on human spleen.
Panel B: anti-CD8 alpha stained on cytotoxic T cells.
Panel C: anti-CD4 stained on T helper cells.
Panel D: anti-CD19 stained on B cells.
Sections were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining: in the order of ab245118 for 30 mins, then ab238798 and ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.DAPI was used as a nuclear counterstain.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. -
Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human tonsil tissue labeling CD8 alpha with ab245118 at 1/500 dilution, CD4 with ab238798 at 1/500, and CD19 with ab237772 at 1/5000 dilution.
Panel A: merged staining of anti-CD8 alpha (magenta; Opal™690), anti-CD4 (green; Opal™520) and anti-CD19 (red; Opal™570) on human tonsil.
Panel B: anti-CD8 alpha stained on cytotoxic T cells.
Panel C: anti-CD4 stained on T helper cells.
Panel D: anti-CD19 stained on B cells.
Sections were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining: in the order of ab245118 for 30 mins, then ab238798 and ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.DAPI was used as a nuclear counterstain.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. -
Flow cytometric analysis of human peripheral blood mononuclear cells (PBMC) labeling CD8 alpha with ab245118 at 1/400 (right panel) compared with a Rabbit monoclonal IgG (ab172730) (left panel). Goat anti rabbit IgG (Dylight® 488, ab98462) at 1/2000 dilution was used as the secondary antibody.
Cells were stained with rabbit IgG (Left) or anti-CD8 alpha RabMab (Right). Then stained with Alexa Fluor® 647-conjugated anti-CD4. The expressions of CD4 and CD8 alpha are mutually exclusive.
Gated on viable lymphocytes.
-
CD8 alpha was immunoprecipitated from 0.35 mg human thymus lysate with ab245118 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab245118 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: Human thymus lysate 10 µg (Input).
Lane 2: ab245118 IP in human thymus lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab245118 in human thymus lysate.Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 30 seconds.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (4)
ab245118 has been referenced in 4 publications.
- Sun J et al. Potential correlation of allograft infiltrating group 2 innate lymphoid cells with acute rejection after liver transplantation. Front Immunol 13:953240 (2022). PubMed: 35967423
- Lu M et al. Pathologic responses to neoadjuvant chemoimmunotherapy in primary limited-stage small-cell lung cancer. Thorac Cancer 13:3208-3216 (2022). PubMed: 36208136
- Li Q et al. PRDM1/BLIMP1 induces cancer immune evasion by modulating the USP22-SPI1-PD-L1 axis in hepatocellular carcinoma cells. Nat Commun 13:7677 (2022). PubMed: 36509766
- Li S et al. A risk signature with inflammatory and immune cells infiltration predicts survival and efficiency of chemotherapy in gastric cancer. Int Immunopharmacol 96:107589 (2021). PubMed: 34162126