Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175] (ab208027)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20175] to hnRNP A1 (citrulline R88 + R92)
  • Suitable for: Flow Cyt, IP, WB, Dot blot
  • Reacts with: Human

Overview

  • Product name

    Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175]
    See all hnRNP A1 primary antibodies

  • Description

    Rabbit monoclonal [EPR20175] to hnRNP A1 (citrulline R88 + R92)

  • Host species

    Rabbit

  • Specificity

    This antibody cross reacts with hnRNP A1L2 (citrulline R88/92), hnRNP A0 (citrulline R81/85) and hnRNP A2B1 (citrulline R95/99).

  • Tested Applications & Species

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

  • Immunogen

    This product was produced with the following immunogens:
    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

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

  • Positive control

    • WB: HEK-293T transfected with PADI4 (WT) and HEK-293T transfected with PADI2 (WT) whole cell lysates treated with CaCl2 and Ionomycin. Flow Cyt: HEK-293T transfected with PADI4 (WT) cells treated with CaCl2 and Ionomycin. IP: HEK-293T transfected with PADI4 (WT) cell lysate treated with CaCl2 and Ionomycin.

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

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR20175

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175] (ab208027)
    Western blot - Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175] (ab208027)
    All lanes : Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175] (ab208027) at 1/5000 dilution

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with empty vector (control) then treated with 10 mM CaCl2 and 10 µM Ionomycin for 2 hours, whole cell lysate
    Lane 2 : HEK-293T transfected with a PADI4 expression vector then treated with 10 mM CaCl2 and 10 µM Ionomycin for 2 hours, whole cell lysate
    Lane 3 : HEK-293T transfected with an empty vector (control) then treated with 10 mM CaCl2 and 10 µM Ionomycin for 2 hours, whole cell lysate
    Lane 4 : HEK-293Ttransfected with a PADI2 expression vector then treated with 10 mM CaCl2 and 10 µM Ionomycin for 2 hours, whole cell lysate

    Lysates/proteins at 20 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: 31,37,39 kDa
    why is the actual band size different from the predicted?



    hnRNP A1(Uniprot P09651), isoform A1-A (34kD) is twenty times more abundant than isoform A1-B (39kD). The molecular weights of hnRNP A1L2 (Q32P51), hnRNP A0 (Q13151) and hnRNP A2B1 (P22626) are 34kD, 31kD and 37kD respectively.

    PAD enzymes (including PADI2 and PADI4) hydrolyze arginine to form citrulline, Ca2+ and ionomycin are also required for this catalytic reaction (PMID: 26360112).

    Blocking/Dilution buffer: 5% NFDM/TBST

    Exposure times: Lane 1-2: 1 second; Lane 3-4: 2 seconds.

  • Dot Blot - Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175] (ab208027)
    Dot Blot - Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175] (ab208027)

    Dot Blot analysis of hnRNP A1 (citrulline R88 + R92) labeled with ab208027 at 1/1000 dilution.

    Lane 1: hnRNP A1 (citrulline R88/92) peptide. 

    Lane 2: hnRNP A3 (citrulline R109/113) peptide.

    Lane 3: hnRNP A0 (citrulline R81/85) peptide.

    Lane 4: hnRNP A2B1 (citrulline R95/99) peptide.

    Lane 5: CCDC51 (citrulline R142) peptide.

    Lane 6: hnRNP A1 non-citrulline peptide.

    Lane 7: hnRNP A1 (citrulline R88) peptide.

    Lane 8: hnRNP A1 (citrulline R92) peptide.

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

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 30 seconds.

  • Flow Cytometry - Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175] (ab208027)
    Flow Cytometry - Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175] (ab208027)

    Flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with PADI4 expression vector, then treated with 10 mM CaCl2 and 10 μM Ionomycin for 2 hours, labeling hnRNP A1 with ab208027 at 1/5000 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079) at 1/2000 dilution was used as the secondary antibody.

    The CaCl2 and Ionomycin treated GFP positive cell population gives a positive signal. (Right panel, Q2).

  • Immunoprecipitation - Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175] (ab208027)
    Immunoprecipitation - Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175] (ab208027)

    hnRNP A1 was immunoprecipitated from 0.35 mg HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) with ab208027 at 1/30 dilution, whole cell lysate. Western blot was performed from the immunoprecipitate using ab208027 at 1/10000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HEK-293T transfected with PADI4 expression vector, then treated with 10 mM CaCl2 and 10μM Ionomycin for 2 hours whole cell lysate 10 μg (Input).

    Lane 2: ab208027 IP in HEK-293T transfected with PADI4 expression vector, then treated with 10 mM CaCl2 and 10 μM Ionomycin for 2 hours whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab208027 in HEK-293T transfected with PADI4 expression vector, then treated with 10 mM CaCl2 and 10 μM Ionomycin for 2 hours whole cell lysate.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 1 second.

  • Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175] (ab208027)
    Anti-hnRNP A1 (citrulline R88 + R92) antibody [EPR20175] (ab208027)

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