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RecombinantRabMAb

Recombinant Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)

Submit a review Submit a question References (1)
Western blot - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
  • Immunocytochemistry/ Immunofluorescence - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
  • Flow Cytometry - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
  • Western blot - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
  • Immunoprecipitation - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
  • Immunocytochemistry/ Immunofluorescence - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
  • Flow Cytometry - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
  • Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22172] to HLA Class 1 ABC
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-HLA Class 1 ABC antibody [EPR22172]
    See all HLA Class 1 ABC primary antibodies

  • Description

    Rabbit monoclonal [EPR22172] to HLA Class 1 ABC

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • ICC/IF: THP-1 and Raji cells. Flow Cyt: Raji and THP-1 cells. IHC-P: Human colonic carcinoma, stomach and liver tissue. WB: Raji, THP-1, HL-60, Jurkat, HepG2, TF-1 and Ramos whole cell lysate. Human tonsil. Recombinant human HLA-A protein (aa24-aa365). Recombinant human HLA-C protein (aa25-aa308). IP: HLAB in THP-1 whole cell lysate.

  • 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
    For more information see here.

    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

  • 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

    EPR22172

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab225636 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 40 kDa (predicted molecular weight: 40 kDa).
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/250.
Flow Cyt
1/600.
IP
1/30.
Notes
WB
1/1000. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/250.
Flow Cyt
1/600.
IP
1/30.

Target

  • Relevance

    HLA-ABC antigen is present on most nucleated cells of humans and Old World monkeys. HLA CLass I is involved in the presentation of foreign antigens to the immune system.

  • Cellular localization

    Cell Membrane; Single-pass type I membrane protein

  • Database links

    • Alternative names

      • Antigen Presenting Molecule antibody
      • Cw*1701 antibody
      • D6S204 antibody
      • FLJ27082 antibody
      • HLA A antibody
      • HLA B antibody
      • HLA C antibody
      • HLA class 1 A antibody
      • HLA class 1 B antibody
      • HLA class 1 C antibody
      • HLA JY3 antibody
      • HLAA antibody
      • HLAB antibody
      • HLAC antibody
      • HLC C antibody
      • Human Leucocyte Antigen C antibody
      • Major Histocompatibility Complex Class I A antibody
      • Major Histocompatibility Complex Class I B antibody
      • Major Histocompatibility Complex Class I C antibody
      • Major histocompatibility complex, class I, A + B + C antibody
      • MHC class I antigen A*1 antibody
      • MHC Class I Antigen antibody
      • MHC class I antigen B*7 antibody
      • MHC class I antigen Cw*1 antibody
      • MHC class I HLA A antibody
      • MHC class I HLA B antibody
      • MHC class I HLA C antibody
      • MHC HLA ABC antibody
      • PSORS1 antibody
      see all

    Images

    • Western blot - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      Western blot - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      All lanes : Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636) at 1/1000 dilution

      Lane 1 : Recombinant human HLA-A protein (aa24-aa365) 10ng
      Lane 3 : Recombinant human HLA-C protein (aa25-aa308) 10ng

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

      Predicted band size: 40 kDa



      This antibody has cross reaction with human HLA-A and HLA-C.# Recombinant human HLA-A protein was purchased from abnova company (H00003105-P01).

      Recombinant human HLA-C protein was purchased from novusbio company (NBP2-23210).

    • Immunocytochemistry/ Immunofluorescence - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      Immunocytochemistry/ Immunofluorescence - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)

      Immunofluorescent analysis of 100% methanol-fixed THP-1 (human monocytic leukemia cell line) cells labeling HLAB with ab225636 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in THP-1 cells. The nuclear counterstain is DAPI (blue).
      Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).
      The negative control is the secondary antibody only.

      Methanol is recommended for fixation as weak signal was observed using 4% PFA.

    • Flow Cytometry - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      Flow Cytometry - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)

      Flow cytometric analysis of Raji (human Burkitt's lymphoma cell line) cells labeling HLAB with ab225636 at 1/600 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti-rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

      Gated on viable cells.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)

      Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling HLAB with ab225636 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining on lymph nodule of human stomach (PMID:25294906) is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Western blot - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      Western blot - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      All lanes : Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636) at 1/1000 dilution

      Lane 1 : Raji (human Burkitt's lymphoma cell line), whole cell lysate
      Lane 2 : THP-1 (human monocytic leukemia cell line), whole cell lysate
      Lane 3 : HL-60 (human promyelocytic leukemia cell line), whole cell lysate
      Lane 4 : Jurkat (human T cell leukemia cell line from peripheral blood), whole cell lysate
      Lane 5 : HepG2 (human liver hepatocellular carcinoma cell line), whole cell lysate
      Lane 6 : TF-1 (human bone marrow erythroleukemia cell line), whole cell lysate
      Lane 7 : Ramos (human Burkitt's lymphoma cell line), whole cell lysate
      Lane 8 : Human tonsil

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

      Predicted band size: 40 kDa
      Observed band size: 40 kDa


      Exposure time: 26 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Weak reactive bands of undetermined proteins are observed in addition to the target band.

       

    • Immunoprecipitation - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      Immunoprecipitation - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)

      HLAB was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia cell line) whole cell lysate with ab225636 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab225636 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
      Lane 1: THP-1 whole cell lysate 10 µg (Input).
      Lane 2: ab225636 IP in THP-1 whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab225636 in THP-1 whole cell lysate.
      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 3 seconds.

    • Immunocytochemistry/ Immunofluorescence - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      Immunocytochemistry/ Immunofluorescence - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)

      Immunofluorescent analysis of 100% methanol-fixed Raji (human Burkitt's lymphoma cell line) cells labeling HLAB with ab225636 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Raji cells. The nuclear counterstain is DAPI (blue).
      Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).
      The negative control is the secondary antibody only.

      Methanol is recommended for fixation as weak signal was observed using 4% PFA.

    • Flow Cytometry - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      Flow Cytometry - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)

      Flow cytometric analysis of THP-1 (human monocytic leukemia cell line) cell line labeling HLAB with ab225636 at 1/600 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

      Gated on viable cells.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)

      Immunohistochemical analysis of paraffin-embedded human liver tissue labeling HLAB with ab225636 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on hepatic sinus endothelium of human liver (PMID: 26963506) is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)

      Immunohistochemical analysis of paraffin-embedded human colonic carcinoma tissue labeling HLAB with ab225636 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining on tumor cells of human colonic carcinoma (PMID: 26310568, PMID: 17014712) is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
      Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)

    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.

    Click here to view the general protocols

    References (1)

    Publishing research using ab225636? Please let us know so that we can cite the reference in this datasheet.

    ab225636 has been referenced in 1 publication.

    • Shen R  et al. Influence of oncogenic mutations and tumor microenvironment alterations on extranodal invasion in diffuse large B-cell lymphoma. Clin Transl Med 10:e221 (2020). PubMed: 33252851

    Customer reviews and Q&As

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