Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-BAFF antibody [EPR19704] (ab203791)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19704] to BAFF
  • Suitable for: ICC/IF, IP, WB
  • Reacts with: Human

Overview

  • Product name

    Anti-BAFF antibody [EPR19704]
    See all BAFF primary antibodies

  • Description

    Rabbit monoclonal [EPR19704] to BAFF

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    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: HEK-293 transfected with a GFP-Myc tagged BAFF vector whole cell lysate; U937 whole cell lysate. ICC/IF: 293T cells transfected with a GFP-tagged BAFF expression construct. IP: U937 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

Images

  • Western blot - Anti-BAFF antibody [EPR19704] (ab203791)
    Western blot - Anti-BAFF antibody [EPR19704] (ab203791)
    All lanes : Anti-BAFF antibody [EPR19704] (ab203791) at 1/5000 dilution

    Lane 1 : Empty vector with GFP-Myc tag (vector control) transfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) transfected with a GFP-Myc tagged BAFF vector whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 31 kDa
    Observed band size: 57 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

     

  • Western blot - Anti-BAFF antibody [EPR19704] (ab203791)
    Western blot - Anti-BAFF antibody [EPR19704] (ab203791)
    Anti-BAFF antibody [EPR19704] (ab203791) at 1/1000 dilution + U937 (Human histiocytic lymphoma cell line) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 31 kDa
    Observed band size: 17,29,31 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Two isoforms have been described in literature (PMID: 12867412).

  • Immunocytochemistry/ Immunofluorescence - Anti-BAFF antibody [EPR19704] (ab203791)
    Immunocytochemistry/ Immunofluorescence - Anti-BAFF antibody [EPR19704] (ab203791)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 293T (Human epithelial cell line from embryonic kidney) cells labeling BAFF with ab203791 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 647) (ab150079) secondary antibody at 1/1000 dilution.

    Confocal image showing positive staining on 293T cells transfected with a GFP-tagged BAFF expression construct.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150079 at 1/1000 dilution.

    ICC/IF was also performed on a few cell lines including U937, no signal was detected.

  • Immunoprecipitation - Anti-BAFF antibody [EPR19704] (ab203791)
    Immunoprecipitation - Anti-BAFF antibody [EPR19704] (ab203791)

    BAFF was immunoprecipitated from 0.35 mg of U937 (Human histiocytic lymphoma cell line) whole cell lysate with ab203791 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab203791 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: U937 whole cell lysate, 10µg (Input).

    Lane 2: ab203791 IP in U937 whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab203791 in U937 whole cell lysate.

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

    Exposure time: 30 seconds.

  • Anti-BAFF antibody [EPR19704] (ab203791)
    Anti-BAFF antibody [EPR19704] (ab203791)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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