Recombinant
RabMAb

Recombinant Anti-HLA A antibody [EP1395Y] (ab52922)

Overview

  • Product name
    Anti-HLA A antibody [EP1395Y]
    See all HLA A primary antibodies
  • Description
    Rabbit monoclonal [EP1395Y] to HLA A
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, IHC - Wholemount, In-Cell ELISA, WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Synthetic peptide within Human HLA A aa 50-150. The exact sequence is proprietary.

  • Positive control
    • IHC-P: Human tonsil tissue; Zebrafish xenograft. ICC/IF: MCF7 and Raji cells. WB: THP-1, A549, HL-60 and Raji cell lysate. IP: THP-1 and A549 cell lysate. Flow Cyt: Raji cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EP1395Y
  • Isotype
    IgG
  • Research areas

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 IHC antigen retrieval protocols.

ICC/IF 1/100.

For unpurified use at 1/250 - 1/500.

Target

  • Relevance
    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.
  • Database links
  • Alternative names
    • 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

Images

  • 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).

  • 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
    All lanes : 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.

  • 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.
  • 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 Detection Reagent (ab131366) (1/10,000) was used for detection. 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.

  • 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.

  • 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

  • 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.

  • 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.

  • 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 Detection Reagent (ab131366) (1/10,000) was used for detection. 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

This product has been referenced in:
  • El-Khoury R  et al. Divergent Features of Mitochondrial Deficiencies in LGMD2A Associated With Novel Calpain-3 Mutations. J Neuropathol Exp Neurol 78:88-98 (2019). Read more (PubMed: 30500922) »
  • Puig P  et al. Multiple immunofluorescence assay identifies upregulation of Active ß-catenin in prostate cancer. BMC Res Notes 12:68 (2019). Read more (PubMed: 30700322) »
See all 36 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA pH 9
Permeabilization
No
Specification
Tonsil
Blocking step
Dako Protein Block X0909 as blocking agent for 15 minute(s) · Concentration: 0.25% · Temperature: 25°C
Fixative
Formaldehyde

Mrs. Amanda Burnette

Verified customer

Submitted Dec 20 2018

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

Dr. Enrico Radaelli

Verified customer

Submitted 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

Abcam user community

Verified customer

Submitted 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

Abcam user community

Verified customer

Submitted 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

Abcam user community

Verified customer

Submitted 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

Dr. Rita Fior

Verified customer

Submitted Jul 14 2014

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

Dr. Rita Fior

Verified customer

Submitted 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

Abcam user community

Verified customer

Submitted Oct 23 2013

Question
Answer

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 93%, so dass es sehr wahrscheinlich ist, dass der Antikörper kreuzreagiert.

Falls Sie dies selbst testen möchten, kann ich Ihnen zur Zeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat zusenden. Der Rabatt würde den ganzen Wert von ab52922abdecken, und gegen eine erneute Bestellung eines weiteren primären Antikörpers von uns verrechnet werden.

Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

1.) Bestätigen Sie mir bitte, dass Sie ab52922am Hund testen möchten.

2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes.

3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

4.) Testen Sie den Antikörper am Hund.

5.) Teilen Sie uns das Ergebnis Ihres Tests durch ein Abreview mit und notieren Sie den Discount Code in dem Feld "additional notes"
Unter der folgenden URL können Sie mehr über unser Abreview System erfahren:
https://www.abcam.com/abreviews

6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv, und Sie können einen anderen primären Antikörper zu dem gleichen Preis wie ab52922bei uns bestellen (halten Sie bei der Bestellung bitte den Rabattcode und die Bestellnummer von ab52922bereit). Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist.

Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
https://www.abcam.com/collaborationdiscount

Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

Read More

Answer

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 in IHC-P.

Generell garantieren wir, dass unsere Antikörper wie auf dem Datenblatt beschrieben funktionieren. Dies schließt jedoch eine spezifische Antigendemaskierungsmethode nicht mit ein und wir erwarten von unseren Anwendern, dass Protokoll und Antigendemaskierungsmethode entsprechend optimiert werden.

Ich habe auch noch einige Informationen über EDTA pH 9 Antigendemaskierungen angehängt. Ich nehme an, sie versuchen eine Doppelfärbung zu optimieren? Vielleicht wäre EDTA -pH9 die richtige Wahl?

Es tut mir leid, dass ich Ihnen in diesem Fall nicht weiterhelfen konnte.

Ich wünsche Ihnen viel Erfolg bei der Suche.

Read More

1-10 of 19 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up