Anti-HLA A antibody [EP1395Y] (ab52922)
- Datasheet
- Specific References (22)
- Protocols
Overview
- Product nameAnti-HLA A antibody [EP1395Y]
See all HLA A primary antibodies - DescriptionRabbit monoclonal [EP1395Y] to HLA A
- Tested applicationsSuitable for: IHC-Fr, IHC - Wholemount, In-Cell ELISA, WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
- Species reactivityReacts with: Rat, Human
- Immunogen
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human HLA A aa 50-150.
- Positive control
- Raji cell lysate, human tonsil.
- General notes
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Alternative versions available:
Anti-HLA A antibody (Alexa Fluor® 488) [EP1395Y] (ab199780)
Anti-HLA A antibody (Alexa Fluor® 647) [EP1395Y] (ab199837)
Anti-HLA A antibody (HRP) [EP1395Y] (ab199555)
Anti-HLA A antibody (Alexa Fluor® 594) [EP1395Y] (ab207872)
Properties
- FormLiquid
- Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
- Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... - PurityProtein A purified
- ClonalityMonoclonal
- Clone numberEP1395Y
- IsotypeIgG
- Research areas
Associated products
- Compatible Secondaries
- Positive Controls
- Related Products
Applications
Our Abpromise guarantee covers the use of ab52922 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-Fr | Use at an assay dependent concentration. | |
| IHC - Wholemount | Use at an assay dependent concentration. | |
| In-Cell ELISA | Use at an assay dependent concentration. | |
| WB | 1/2000. Predicted molecular weight: 41 kDa. For unpurified use at 1/10000 - 1/50000. |
|
| IP | 1/20. For unpurified use at 1/30. |
|
| Flow Cyt | 1/10 - 1/100. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. | |
| IHC-P | 1/100. For unpurified use at 1/250 - 1/500. See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol). |
|
| ICC/IF | 1/100. For unpurified use at 1/250 - 1/500. |
Target
- RelevanceHLA-A belongs to the HLA class I heavy chain paralogues. This class I molecule is a heterodimer consisting of a heavy chain and a light chain (beta-2 microglobulin). The heavy chain is anchored in the membrane. Class I molecules play a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen. They are expressed in nearly all cells. The heavy chain is approximately 45 kDa and its gene contains 8 exons. Exon 1 encodes the leader peptide, exons 2 and 3 encode the alpha1 and alpha2 domains, which both bind the peptide, exon 4 encodes the alpha3 domain, exon 5 encodes the transmembrane region, and exons 6 and 7 encode the cytoplasmic tail. Polymorphisms within exon 2 and exon 3 are responsible for the peptide binding specificity of each class one molecule. Typing for these polymorphisms is routinely done for bone marrow and kidney transplantation. Hundreds of HLA-A alleles have been described.
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Database links
- Entrez Gene: 3105 Human
- Omim: 142800 Human
- SwissProt: P01891 Human
- SwissProt: P01892 Human
- SwissProt: P10314 Human
- SwissProt: P10316 Human
- SwissProt: P13746 Human
- SwissProt: P16188 Human
see all -
Alternative names
- Antigen presenting molecule antibody
- HLA class I histocompatibility antigen, A 1 alpha chain antibody
- HLAA antibody
see all
Anti-HLA A antibody [EP1395Y] images
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![IHC - Wholemount - Anti-HLA A [EP1395Y] antibody (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/ab52922-229806-ab52922ihcp.jpg)
IHC - Wholemount - Anti-HLA A [EP1395Y] antibody (ab52922)This image is courtesy of an Abreview submitted by Rita FiorIHC - Wholemount of human zebrafish xenograft labelling HLA A with ab52922. Sample was incubated with primary antibody (1/100) for 1 hours at 4°C. An Alexa Fluor® 647-conjugated goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.
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![Immunocytochemistry/ Immunofluorescence - Anti-HLA A antibody [EP1395Y] (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/MHC-class-I-Primary-antibodies-ab52922-4.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-HLA A antibody [EP1395Y] (ab52922)ICC/IF image of unpurified ab52922 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52922, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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![Western blot - Anti-HLA A antibody [EP1395Y] (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/ab52922-14-ab52922WB2.jpg)
Western blot - Anti-HLA A antibody [EP1395Y] (ab52922)All lanes : Anti-HLA A antibody [EP1395Y] (ab52922) at 1/5000 dilution (purified)
Lane 1 : THP-1 cell lysate at 20 µg
Lane 2 : A549 cell lysate at 1/20 dilution
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size : 41 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST. -
![Western blot - Anti-HLA A antibody [EP1395Y] (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/ab52922-13-ab52922WB1.jpg)
Western blot - Anti-HLA A antibody [EP1395Y] (ab52922)Anti-HLA A antibody [EP1395Y] (ab52922) at 1/2000 dilution (purified) + HL-60 cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size : 41 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST. -
![Western blot - Anti-HLA A [EP1395Y] antibody (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/ab52922wb.jpg)
Western blot - Anti-HLA A [EP1395Y] antibody (ab52922)Anti-HLA A antibody [EP1395Y] (ab52922) at 1/10000 dilution + Raji cell lysate at 10 µg
Secondary
Goat anti rabbit IgG HRP conjugated at 1/2000 dilution
Predicted band size : 41 kDa
Observed band size : 41 kDa -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA A antibody [EP1395Y] (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/ab52922-8-ab52922IHC1.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA A antibody [EP1395Y] (ab52922)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling HLA A with purified ab52922 at 1/100. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA A antibody [EP1395Y] (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/ab52922-210120-ab52922ihcp.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA A antibody [EP1395Y] (ab52922)This image is courtesy of an anonymous Abreviewab52922 staining HLA A in Human tonsil tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% H2O2 for 10 minutes at 25°C; antigen retrieval was by heat mediation in a citrate buffer, pH 6.0 . Samples were incubated with primary antibody (1/3000) for 20 minutes at 25°C. An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA A antibody [EP1395Y] (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/ab52922-249497-new-ab52922.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA A antibody [EP1395Y] (ab52922)Ab52922 at 1/250 dilution staining human tonsil; paraffin embedded.
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![Immunocytochemistry/ Immunofluorescence - Anti-HLA A antibody [EP1395Y] (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/ab52922-9-ab52922ICCIF1.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-HLA A antibody [EP1395Y] (ab52922)Immunocytochemistry/Immunofluorescence analysis of Raji (human Burkitt's lymphoma) cells labelling HLA A with purified ab52922 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
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![Flow Cytometry - Anti-HLA A antibody [EP1395Y] (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/ab52922-10-ab52922FC1.jpg)
Flow Cytometry - Anti-HLA A antibody [EP1395Y] (ab52922)Flow Cytometry analysis of Raji cells labelling HLA A with purified ab52922 at 1/40 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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![Flow Cytometry - Anti-HLA A [EP1395Y] antibody (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/MHC-class-I-Primary-antibodies-ab52922-8.jpg)
Flow Cytometry - Anti-HLA A [EP1395Y] antibody (ab52922)Overlay histogram showing Raji cells stained with ab52922 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52922, 1/100) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Raji cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions. -
![Immunoprecipitation - Anti-HLA A antibody [EP1395Y] (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/ab52922-11-ab52922IP1.jpg)
Immunoprecipitation - Anti-HLA A antibody [EP1395Y] (ab52922)ab52922 (purified) at 1/20 immunoprecipitating HLA A in THP-1 whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP secondary antibody (ab131366) (1/10,000) was used as the secondary antibody. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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![Immunoprecipitation - Anti-HLA A antibody [EP1395Y] (ab52922)](http://a.abcam.com/ps/products/52/ab52922/Images/ab52922-12-ab52922IP2.jpg)
Immunoprecipitation - Anti-HLA A antibody [EP1395Y] (ab52922)ab52922 (purified) at 1/20 immunoprecipitating HLA A in A549 whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP secondary antibody (ab131366) (1/10,000) was used as the secondary antibody. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
References for Anti-HLA A antibody [EP1395Y] (ab52922)
This product has been referenced in:
- Ritz D et al. High-sensitivity HLA class I peptidome analysis enables a precise definition of peptide motifs and the identification of peptides from cell lines and patients' sera. Proteomics N/A:N/A (2016). WB ; Human . Read more (PubMed: 26992070) »
- Grau JJ et al. Enrichment of Cells with Cancer Stem Cell-Like Markers in Relapses of Chemoresistant Patients with Locally Advanced Head and Neck Squamous Cell Carcinoma. Oncology 90:267-72 (2016). IHC ; Human . Read more (PubMed: 27077749) »
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