Recombinant Anti-CD8 alpha antibody [EPR21769] (ab217344)
![Flow Cytometry - Anti-CD8 alpha antibody [EPR21769] (ab217344)](https://www.abcam.com/ps/products/217/ab217344/Images/ab217344-297302-anti-cd8-alpha-antibody-epr21769-flow-cytometry.jpg "Flow Cytometry - 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... -
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 |
|---|---|---|
| WB |
1/1000. Detects a band of approximately 34-38 kDa (predicted molecular weight: 27 kDa).
|
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| IHC-Fr |
1/500.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
| IHC-P | (2) |
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 |
|---|
|
WB
1/1000. Detects a band of approximately 34-38 kDa (predicted molecular weight: 27 kDa). |
|
IHC-Fr
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
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. |
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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 Cytometry - Anti-CD8 alpha antibody [EPR21769] (ab217344)
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)](https://www.abcam.com/ps/products/217/ab217344/Images/ab217344-297300-anti-cd8-alpha-antibody-epr21769-immunohistochemistry.jpg)
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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![Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)](https://www.abcam.com/ps/products/217/ab217344/Images/ab217344-297298-anti-cd8-alpha-antibody-epr21769-immunohistochemistry.jpg)
Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)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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![Immunoprecipitation - Anti-CD8 alpha antibody [EPR21769] (ab217344)](https://www.abcam.com/ps/products/217/ab217344/Images/ab217344-297303-anti-cd8-alpha-antibody-epr21769-immunoprecipitation.jpg)
Immunoprecipitation - Anti-CD8 alpha antibody [EPR21769] (ab217344)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)](https://www.abcam.com/ps/products/217/ab217344/Images/ab217344-297301-anti-cd8-alpha-antibody-epr21769-immunohistochemistry.jpg)
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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![Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)](https://www.abcam.com/ps/products/217/ab217344/Images/ab217344-297299-anti-cd8-alpha-antibody-epr21769-immunohistochemistry.jpg)
Immunohistochemistry (Frozen sections) - Anti-CD8 alpha antibody [EPR21769] (ab217344)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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![Western blot - Anti-CD8 alpha antibody [EPR21769] (ab217344)](https://www.abcam.com/ps/products/217/ab217344/Images/ab217344-297297-anti-cd8-alpha-antibody-epr21769-western-blot.jpg)
Western blot - Anti-CD8 alpha antibody [EPR21769] (ab217344)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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![Anti-CD8 alpha antibody [EPR21769] (ab217344)](https://www.abcam.com/ps/products/217/ab217344/Images/ab217344-8-benefits-of-recombinant-antibodies.png)
Anti-CD8 alpha antibody [EPR21769] (ab217344)
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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Datasheet download
Certificate of Compliance
References (39)
ab217344 has been referenced in 39 publications.
- Huang Z et al. Nanoscale coordination polymers induce immunogenic cell death by amplifying radiation therapy mediated oxidative stress. Nat Commun 12:145 (2021). PubMed: 33420008
- Yoneyama T et al. Tumor vasculature-targeted 10B delivery by an Annexin A1-binding peptide boosts effects of boron neutron capture therapy. BMC Cancer 21:72 (2021). PubMed: 33446132
- Upadhyaya P et al. Anticancer immunity induced by a synthetic tumor-targeted CD137 agonist. J Immunother Cancer 9:N/A (2021). PubMed: 33500260
- Cheng JN et al. Radiation-induced eosinophils improve cytotoxic T lymphocyte recruitment and response to immunotherapy. Sci Adv 7:N/A (2021). PubMed: 33514544
- Li L et al. Laminin ?2-mediating T cell exclusion attenuates response to anti-PD-1 therapy. Sci Adv 7:N/A (2021). PubMed: 33536206
