Recombinant Anti-CD8 alpha antibody [EPR21769] (ab217344)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21769] to CD8 alpha
- Suitable for: WB, IHC-Fr, IHC-P, Flow Cyt, IP
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-CD8 alpha antibody [EPR21769]
See all CD8 alpha primary antibodies -
Description
Rabbit monoclonal [EPR21769] to CD8 alpha -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Fr, IHC-P, Flow Cyt, IPmore details -
Species reactivity
Reacts with: Mouse -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse thymus, spleen and lymph node lysates. IHC-P: Mouse spleen and colon tissues. IHC-Fr: Mouse spleen and thymus tissues. Flow Cyt: Mouse primary splenocytes. IP: Mouse thymus lysate.
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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
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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, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21769 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Anti-CD8 alpha antibody [EPR21769] - BSA and Azide free (ab230156)
- Alexa Fluor® 488 Anti-CD8 alpha antibody [EPR21769] (ab237364)
- Alexa Fluor® 647 Anti-CD8 alpha antibody [EPR21769] (ab237365)
- FITC Anti-CD8 alpha antibody [EPR21769] (ab237367)
- APC Anti-CD8 alpha antibody [EPR21769] (ab237368)
- Alexa Fluor® 555 Anti-CD8 alpha antibody [EPR21769] (ab280863)
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab217344 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 |
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WB |
1/1000. Detects a band of approximately 34-38 kDa (predicted molecular weight: 27 kDa).
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IHC-Fr |
1/500.
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IHC-P | (4) |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Background staining was observed in mouse testis samples. |
Flow Cyt |
1/500.
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IP |
1/30.
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Notes |
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WB
1/1000. Detects a band of approximately 34-38 kDa (predicted molecular weight: 27 kDa). |
IHC-Fr
1/500. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Background staining was observed in mouse testis samples. |
Flow Cyt
1/500. |
IP
1/30. |
Target
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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
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Database links
- Entrez Gene: 12525 Mouse
- SwissProt: P01731 Mouse
- Unigene: 1858 Mouse
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Alternative names
- alpha polypeptide (p32) antibody
- CD_antigen=CD8a antibody
- CD8 antibody
see all
Images
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Flow cytometric analysis of mouse primary splenocytes labeling CD8 alpha with ab217344 at 1/500 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Cells were surface stained with CD4-Alexa Fluor® 647, then stained with rabbit IgG (Left) / ab217344 (Right) separately. CD4 and CD8 alpha are mutually exclusive expressed in mouse spleen. Gated on total viable cells.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD8 alpha with ab217344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on the white pulp of mouse spleen (PMID: 23482450; PMID: 25826597). Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen tissue labeling CD8 alpha with ab217344 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive membrane staining on mouse spleen (PMID: 25616911).
The nuclear counter stain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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CD8 alpha was immunoprecipitated from 0.35 mg of mouse thymus lysate with ab217344 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab217344 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: Mouse thymus lysate 10 µg (Input).
Lane 2: ab217344 IP in mouse thymus lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214344 in mouse thymus lysate.
Exposure time: 5 seconds.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
The two bands are different isoforms that are consistent with the literature (PMID 3085089).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)
Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling CD8 alpha with ab217344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on stromal cells of mouse colon. Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
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Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse thymus tissue labeling CD8 alpha with ab217344 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive membrane staining on mouse thymus tissue section (PMID: 25616911).
The nuclear counter stain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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All lanes : Anti-CD8 alpha antibody [EPR21769] (ab217344) at 1/1000 dilution
Lane 1 : Mouse thymus lysate at 20 µg
Lane 2 : Mouse spleen lysate at 20 µg
Lane 3 : Mouse lymph node lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 27 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?Exposure time : Lane 1: 3 sconds; Lane 2: 40 seconds; Lane 3: 15 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
The two bands are different isoforms that are consistent with the literature (PMID 3085089).
The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)
Tissue Microarrays stained for "Anti-CD8 alpha antibody [EPR21769]” using "ab217344" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab217344 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
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
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SDS download
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Datasheet download
Certificate of Compliance
References (96)
ab217344 has been referenced in 96 publications.
- Hao B et al. Single-cell RNA sequencing analysis revealed cellular and molecular immune profiles in lung squamous cell carcinoma. Transl Oncol 27:101568 (2023). PubMed: 36270103
- Chen T et al. Enhancement of T cell infiltration via tumor-targeted Th9 cell delivery improves the efficacy of antitumor immunotherapy of solid tumors. Bioact Mater 23:508-523 (2023). PubMed: 36514387
- Xiao J et al. UBC9 deficiency enhances immunostimulatory macrophage activation and subsequent antitumor T cell response in prostate cancer. J Clin Invest 133:N/A (2023). PubMed: 36626227
- Gu Z et al. HNRNPC, a predictor of prognosis and immunotherapy response based on bioinformatics analysis, is related to proliferation and invasion of NSCLC cells. Respir Res 23:362 (2022). PubMed: 36536402
- Han SJ et al. Steroid receptor coactivator-3 inhibition generates breast cancer antitumor immune microenvironment. Breast Cancer Res 24:73 (2022). PubMed: 36316775