製品の概要

  • 製品名Anti-HLA A antibody [EP1395Y]
    HLA A 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP1395Y] to HLA A
  • アプリケーション適用あり: IHC-Fr, IHC - Wholemount, In-Cell ELISA, WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • 種交差性
    交差種: Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human HLA A aa 50-150.

  • ポジティブ・コントロール
    • Raji cell lysate, human tonsil.
  • 特記事項

    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)

     

製品の特性

  • 製品の状態Liquid
  • 保存方法Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • バッファーpH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • 精製度Protein A purified
  • ポリ/モノモノクローナル
  • クローン名EP1395Y
  • アイソタイプIgG
  • 研究分野

アプリケーション

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.

アプリ���ーション Abreviews 特記事項
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.

ターゲット情報

  • 関連性HLA-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.
  • 参照データベース
  • 別名
    • Antigen presenting molecule antibody
    • HLA class I histocompatibility antigen, A 1 alpha chain antibody
    • HLAA antibody
    • Leukocyte antigen class I A antibody
    • Major histocompatibility complex, class I, A antibody
    • MHC class I antigen HLA A heavy chain antibody
    see all

Anti-HLA A antibody [EP1395Y] 画像

  • IHC - 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.

    See Abreview

  • 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.

  • 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 kDa

    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • 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 kDa

    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • 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) 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.
  • ab52922 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.

    See Abreview

  • Ab52922 at 1/250 dilution staining human tonsil; paraffin embedded.

  • 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).

  • 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.

  • 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.

  • 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.

  • 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.

Anti-HLA A antibody [EP1395Y] (ab52922) 使用論文

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) »

See all 22 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (human leukemia xenotransplanted into NGS mice)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate pH6
Permeabilization Yes - Tween-20
Specification human leukemia xenotransplanted into NGS mice
Blocking step BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Formaldehyde
Username

enrico radaelli

Verified customer

投稿 Jun 13 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (NSCLC)
Antigen retrieval step None
Permeabilization No
Specification NSCLC
Blocking step Zytomed Systems HRP-Polymer Blocking Reagent as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative HOPE-fixation
Username

Abcam user community

Verified customer

投稿 Oct 23 2015

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (MCF7)
Specification MCF7
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

投稿 Feb 10 2015

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (Raji cell lysate)
Specification Raji cell lysate
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Feb 09 2015

Application ELISA
Sample Human Cell (In Cell ELISA CRC SW620 cell line)
Specification In Cell ELISA CRC SW620 cell line
Type Direct
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Dr. Rita Fior

Verified customer

投稿 Jul 14 2014

Application IHC - Wholemount
Sample Human Embryo (human zebrafish xenograft)
Specification human zebrafish xenograft
Username

Dr. Rita Fior

Verified customer

投稿 Jul 14 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate ph6, 100C for 20 min
Sample Human Tissue sections (tonsil)
Specification tonsil
Permeabilization No
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 Oct 23 2013

Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

Unseres Wissens nach wurde ab52922 Anti-MHC class I antibody [EP1395Y] bisher noch nichtamHundgetestet.

Die Homologie mit dem Hundeprotein beträgt ...

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Vielen Dank für Ihre Anfrage.

Die meisten unserer Antikörper sindmit hitzeinduzierter Antigendemaskierung (HIER)getestet. Leider konnte ich keinen AntiMHC Klasse1 Antikörper finden, der schon einmalmit Proteinase getestet wurde...

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Unfortunately, we do not know if this antibody will cross-react with the human MHC-Class 1 heavy chain.

Searching our catalogue, we do not have any other antibody which would fit your descriptions.

However, on the second look, ab...

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1-10 of 18 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"