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Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23244-12] to Junctional Adhesion Molecule 1/JAM-A
  • Suitable for: ICC/IF, WB, Flow Cyt, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12]
    See all Junctional Adhesion Molecule 1/JAM-A primary antibodies

  • Description

    Rabbit monoclonal [EPR23244-12] to Junctional Adhesion Molecule 1/JAM-A

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: Human lung tissue lysate. HT-29, HUVEC and TF-1 whole cell lysate. ICC/IF: HT-29 cells. Flow Cyt: HT-29 cells, human B lymphocytes and human monocytes. IP: HT-29 whole cell lysate. Human lung tissue 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

Images

  • Western blot - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)
    Western blot - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)
    All lanes : Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948) at 1/1000 dilution

    Lane 1 : Human lung lysate
    Lane 2 : HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
    Lane 3 : THP-1 (human monocytic leukemia monocyte) whole cell lysate
    Lane 4 : HUVEC (human umbilical vein endothelial cell) whole cell lysate
    Lane 5 : MOLT-4 (human lymphoblastic leukemia t lymphoblast) whole cell lysate
    Lane 6 : TF-1 (human erythroleukemia erythroblast) whole cell lysate

    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: 32 kDa
    Observed band size: 35 kDa
    why is the actual band size different from the predicted?



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

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:10698320, 10753840).

    Low expression control: THP-1 and Molt-4 (PMID:10698320).

    Exposure time: Lanes 1-5: 15 seconds; Lane 6:70 seconds.

  • Immunoprecipitation - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)
    Immunoprecipitation - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)

    Junctional Adhesion Molecule 1/JAM-A was immunoprecipitated from 0.35 mg human lung lysate 10µg with ab269948 at 1/30 dilution (2µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab269948 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

    Lane 1: Human lung lysate 10µg.

    Lane 2: ab269948 IP in human lung lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab269948 in human lung lysate.

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

    Exposure time: 15 seconds.

  • Immunoprecipitation - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)
    Immunoprecipitation - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)

    Junctional Adhesion Molecule 1/JAM-A was immunoprecipitated from 0.35 mg HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate 10µg with ab269948 at 1/30 dilution (2µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab269948 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

    Lane 1: HT-29 whole cell lysate 10µg.

    Lane 2: ab269948 IP in HT-29 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab269948 in HT-29 whole cell lysate.

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

    Exposure time: 15 seconds.

     

  • Flow Cytometry - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)
    Flow Cytometry - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)

    Flow cytometric analysis of human B lymphocytes (Left) / human monocytes (Right) cells labeling Junctional Adhesion Molecule 1/JAM-A with ab269948 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control 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 was used as the secondary antibody.

    Human peripheral blood mononuclear cell (PBMC) co-stained with anti-CD19 conjugated to PE-Cy7 and anti-CD14 conjugated to BV510. JAM-A expression on B lymphocytes (CD19+) and monocytes (CD14+) population are shown respectively.

    Gated on viable cells.

  • Flow Cytometry - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)
    Flow Cytometry - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)

    Flow cytometric analysis of HT-29 (Human colorectal adenocarcinoma epithelial cell) cells labeling Junctional Adhesion Molecule 1/JAM-A with ab269948 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control 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 was used as the secondary antibody.

    Gated on viable cells.

  • Immunocytochemistry/ Immunofluorescence - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)
    Immunocytochemistry/ Immunofluorescence - Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT-29 (human colorectal adenocarcinoma epithelial cell) cells labeling Junctional Adhesion Molecule 1/JAM-A with ab269948 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in HT-29 cell line is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  • Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)
    Anti-Junctional Adhesion Molecule 1/JAM-A antibody [EPR23244-12] (ab269948)

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