Recombinant
RabMAb

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

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
    Suitable for: Flow Cyt, IP, WB, Dot blotmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    This product was produced with the following immunogens:
    Synthetic peptide corresponding to Human hnRNP A1 aa 50-150 (citrulline R88 + R92).
    Database link: P09651

    Synthetic peptide within Human hnRNP A1 aa 50-150 (citrulline R88 + R92). The exact sequence is proprietary.
    Database link: P09651

  • 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

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

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
    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol, PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR20175
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab208027 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
Flow Cyt 1/5000.
IP 1/30.
WB 1/1000. Detects a band of approximately 31, 34, 37 kDa (predicted molecular weight: 38 kDa).
Dot blot 1/1000.

Target

  • Function
    Involved in the packaging of pre-mRNA into hnRNP particles, transport of poly(A) mRNA from the nucleus to the cytoplasm and may modulate splice site selection. May play a role in HCV RNA replication.
  • Sequence similarities
    Contains 2 RRM (RNA recognition motif) domains.
  • Post-translational
    modifications
    Arg-194, Arg-206 and Arg-225 are dimethylated, probably to asymmetric dimethylarginine.
    Sumoylated.
  • Cellular localization
    Nucleus. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Shuttles continuously between the nucleus and the cytoplasm along with mRNA. Component of ribonucleosomes. In the course of viral infection, colocalizes with HCV NS5B at speckles in the cytoplasm in a HCV-replication dependent manner.
  • Information by UniProt
  • Database links
  • Alternative names
    • HNRNPA 1 antibody
    • Helix destabilizing protein antibody
    • Helix-destabilizing protein antibody
    • Heterogeneous nuclear ribonucleoprotein A1 antibody
    • Heterogeneous nuclear ribonucleoprotein A1B protein antibody
    • Heterogeneous nuclear ribonucleoprotein B2 protein antibody
    • Heterogeneous nuclear ribonucleoprotein core protein A1 antibody
    • hnRNP A1 antibody
    • hnRNP core protein A1 antibody
    • HNRNPA1 antibody
    • HNRPA1 antibody
    • MGC102835 antibody
    • Nuclear ribonucleoprotein particle A1 protein antibody
    • ROA1_HUMAN antibody
    • Single strand DNA binding protein UP1 antibody
    • Single strand RNA binding protein antibody
    • Single-strand RNA-binding protein antibody
    see all

Images

  • 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.

  • 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.

  • 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).

  • 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 secondary antibody (HRP) (ab131366), was used as secondary antibody 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.

References

ab208027 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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