Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-hnRNP A1 (citrulline R97) antibody [EPR20176] (ab208028)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20176] to hnRNP A1 (citrulline R97)
  • Suitable for: WB, IP, Dot blot
  • Reacts with: Human, Recombinant fragment

Overview

  • Product name

    Anti-hnRNP A1 (citrulline R97) antibody [EPR20176]
    See all hnRNP A1 primary antibodies

  • Description

    Rabbit monoclonal [EPR20176] to hnRNP A1 (citrulline R97)

  • Host species

    Rabbit

  • Specificity

    Based on sequence homology we predict that this antibody may react with hnRNP A1L2.

  • Tested Applications & Species

    Application Species
    Dot
    Recombinant fragment
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK-293T transfected with PADI2 and PADI4, respectively, treated with CaCl2 and Ionomycin, whole cell lysates. IP: HEK-293T transfected with PADI4, treated with CaCl2 and Ionomycin, 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR20176

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-hnRNP A1 (citrulline R97) antibody [EPR20176] (ab208028)
    Western blot - Anti-hnRNP A1 (citrulline R97) antibody [EPR20176] (ab208028)
    Lanes 1-2 & 4 : Anti-hnRNP A1 (citrulline R97) antibody [EPR20176] (ab208028) at 1/5000 dilution
    Lane 3 : Anti-hnRNP A1 (citrulline R97) antibody [EPR20176] (ab208028) at 1/1000 dilution

    Lanes 1 & 3 : Empty vector (vector control) transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen), whole cell lysate
    Lane 2 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate transfected with GFP-tagged PADI2 and treated with 10 mM CaCl2 for 2 hours, followed by 10 uM Ionomycin for 2 hours
    Lane 4 : HEK-293T (human epithelial cell line from embryonic kidney tranformed with large T antigen) whole cell lysate transfected with GFP- tagged PADI4 and treated with 10 mM CaCl2 for 2 hours, followed by 10 uM Ionomycin for 2 hours

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    Predicted band size: 38 kDa
    Observed band size: 34 kDa
    why is the actual band size different from the predicted?


    Exposure time: 4 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    According to Uniprot annotation, isoform A1-A (34kD) is twenty times more abundant than isoform A1-B (39kD).  PADs (including PADI2 and PADI4) are the enzymes that hydrolyze arginine to form citrulline, and this catalytic reaction also needs Ca2+ and ionomycin.  Please refer to PMID: 26360112.

  • Dot Blot - Anti-hnRNP A1 (citrulline R97) antibody [EPR20176] (ab208028)
    Dot Blot - Anti-hnRNP A1 (citrulline R97) antibody [EPR20176] (ab208028)

    Dot blot analysis of hnRNP A1 (citrulline R97) labeled with ab208028 at 1/1000 dilution.

    Lane 1: hnRNP A1 (citrulline R97) peptide.
    Lane 2:hnRNP A1 (citrulline R97) non-citrulline peptide.

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-hnRNP A1 (citrulline R97) antibody [EPR20176] (ab208028)
    Immunoprecipitation - Anti-hnRNP A1 (citrulline R97) antibody [EPR20176] (ab208028)

    hnRNP A1 (citrulline R97) was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GFP tagged PADI4 and treated with 10 mM CaCl2 for 2 hours, then 10 µM Ionomycin for 2 hours whole cell lysate with ab208028 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab208028 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HEK-293T transfected with GFP tagged PADI4 and treated with 10 mM CaCl2 for 2 hours, then 10 µM Ionomycin for 2 hours whole cell lysate 10 µg (Input). 
    Lane 2: ab208028 IP in HEK-293T transfected with GFP tagged PADI4 and treated with 10 mM CaCl2 for 2 hours, then 10 µM Ionomycin for 2 hours whole cell lysate. 
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab208028 in HEK-293T transfected with GFP tagged PADI4 and treated with 10 mM CaCl2 for 2 hours, then 10 µM Ionomycin for 2 hours whole cell lysate.

     

    Exposure time: 10 seconds.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Anti-hnRNP A1 (citrulline R97) antibody [EPR20176] (ab208028)
    Anti-hnRNP A1 (citrulline R97) antibody [EPR20176] (ab208028)

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