Anti-CD8 alpha antibody [OX-8] (ab33786)
Key features and details
- Mouse monoclonal [OX-8] to CD8 alpha
- Suitable for: Flow Cyt, ICC/IF, IHC-P, WB
- Reacts with: Rat
- Isotype: IgG1
Related conjugates and formulations
Overview
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Product name
Anti-CD8 alpha antibody [OX-8]
See all CD8 alpha primary antibodies -
Description
Mouse monoclonal [OX-8] to CD8 alpha -
Host species
Mouse -
Tested applications
Suitable for: Flow Cyt, ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Rat -
Immunogen
Tissue, cells or virus. High molecular weight rat thymocyte glycoproteins
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Positive control
- WB: Rat thymus tissue lysate. IHC-P: Rat Spleen. Flow Cyt: Rat splenocytes. ICC/IF: Rat splenocyte cells.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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: 7.20
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
OX-8 -
Myeloma
NS1 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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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 ab33786 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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Flow Cyt |
1/125000.
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P | (2) |
Use a concentration of 5 µg/ml.
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WB |
Use a concentration of 1 µg/ml. Predicted molecular weight: 26 kDa.
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Notes |
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Flow Cyt
1/125000. |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use a concentration of 5 µg/ml. |
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 26 kDa. |
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: 24930 Rat
- SwissProt: P07725 Rat
- Unigene: 10306 Rat
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Alternative names
- alpha polypeptide (p32) antibody
- CD_antigen=CD8a antibody
- CD8 antibody
see all
Images
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ab33786 staining CD8 alpha in Rat splenocytes cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab33786 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 100% methanol (5 min).
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IHC image of CD8 alpha staining in a section of formalin-fixed paraffin-embedded normal Rat Spleen performed on a Leica BOND™ system using the standard Protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab33923, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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All lanes : Anti-CD8 alpha antibody [OX-8] (ab33786) at 1 µg/ml
Lane 1 : Rat thymus tissue lysate
Lane 2 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 26 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?This blot was produced using a 4-12% Bis-tris under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour using 3% milk before ab33786 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at a 1 μg/ml concentration and 1/20000 dilution respectively. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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ab33786 staining CD8 alpha in rat spleen tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Tissue was fixed with 10% neutral buffered formalin and a heat mediated antigen retrieval step was performed using citrate buffer. Samples were then permeabilized with Triton X-100, blocked using 5% serum for 30 minutes at 25°C and then incubated with ab33786 at a 1/100 dilution for 1 hour at 25°C. The secondary used was a biotin-conjugated rabbit anti-mouse polyclonal, used at a 1/300 dilution. -
Flow cytometry staining of Lewis rat splenocytes with ab33786 (right) or mouse IgG1 kappa; (ab170190) isotype (left). Cells were incubated for 30 min on ice in 1x PBS containing 10 % rat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab33786) or mouse IgG1 kappa; (ab170190) isotype (1x 106 in 100 µl at 0.008 µg/ml) for 30 min on ice.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150177) was used at 1/2000 dilution for 30 min on ice.
The cells were simultaneously stained with CD4.
Acquisition of >30,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on live CD3 positive T cells.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (33)
ab33786 has been referenced in 33 publications.
- Bertho A et al. Evaluation of the Role of the Immune System Response After Minibeam Radiation Therapy. Int J Radiat Oncol Biol Phys 115:426-439 (2023). PubMed: 35985455
- Cao W et al. Transcriptomic analysis of graft liver provides insight into the immune response of rat liver transplantation. Front Immunol 13:947437 (2022). PubMed: 36426354
- Vainio SK et al. Dimethyl fumarate decreases short-term but not long-term inflammation in a focal EAE model of neuroinflammation. EJNMMI Res 12:6 (2022). PubMed: 35107664
- Zhang S et al. Nerve regeneration in rat peripheral nerve allografts: An assessment of the role of endogenous neurotrophic factors in nerve cryopreservation and regeneration. Eur J Neurosci 55:1895-1916 (2022). PubMed: 35332602
- Yang QY et al. Effects of intraperitoneal injection of lipopolysaccharide-induced peripheral inflammation on dopamine neuron damage in rat midbrain. CNS Neurosci Ther 28:1624-1636 (2022). PubMed: 35789066