Overview

  • Product nameAnti-hnRNP A1 antibody [9H10]See all hnRNP A1 primary antibodies ...
  • Description
    Mouse monoclonal [9H10] to hnRNP A1
  • Tested applicationsFlow Cyt, IHC-P, ELISA, WB, IP, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Full length hnRNPA1 native protein (partially purified) from HeLa cells (Human).

Properties

  • FormLiquid
  • Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: 0.1% Sodium Azide
    Constituents: PBS
  • Concentration information loading...
  • PurityProtein A purified
  • Purification notesPurified from tissue culture supernatant.
  • Clonality Monoclonal
  • Clone number9H10
  • MyelomaSp2/0
  • IsotypeIgG2b
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab5832 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
Flow Cyt Use 1µg for 106 cells.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ELISA Use at an assay dependent dilution.
WB Use at an assay dependent dilution. Detects a band of approximately 34 kDa (predicted molecular weight: 38 kDa).
IP Use at an assay dependent dilution. This antibody does not IP the hnRNP complex.
ICC/IF Use at an assay dependent dilution. See Abreview (April 2, 2007)

Target

  • FunctionInvolved 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 similaritiesContains 2 RRM (RNA recognition motif) domains.
  • Post-translational
    modifications
    Arg-194, Arg-206 and Arg-225 are dimethylated, probably to asymmetric dimethylarginine.
    Sumoylated.
  • Cellular localizationNucleus. 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.
  • Target information above from: UniProt accession P09651 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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
    • syncrip antibody
    see all

Anti-hnRNP A1 antibody [9H10] images

  • ab5832 at 1/2000 staining GM10725 cells (Lymphoblastoid cell line). The cells were paraformaldehyde fixed and blocked with BSA prior to incubation with the antibody for 12 hours. An Alexa-Fluor ® 594 conjugated donkey anti-mouse antibody was used as the secondary. Cells were mounted in DAPI and observed using a confocal microscope.

    See Abreview

  • Ab5832 staining human lung. Staining is localized to the nucleus.
    Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system (Dako PT Link), at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer, citrate pH 6.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • All lanes : Anti-hnRNP A1 antibody [9H10] (ab5832) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 38 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 43 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 30 seconds
  • All lanes : Anti-hnRNP A1 antibody [9H10] (ab5832) at 1/1000 dilution

    Lane 1 : Mouse NSC34 whole cell lysate with control siRNA
    Lane 2 : Mouse NSC34 whole cell lysate with hnRNP A1 siRNA

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye® 700DX-conjugated Donkey anti-Mouse IgG at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 38 kDa


    Exposure time : 1 minute

    This image is courtesy of an anonymous Abreview.

    See Abreview

  • Overlay histogram showing Jurkat cells stained with ab5832 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab5832, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • hnRNP A1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to hnRNP A1 (ab5832) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab5832.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 37kDa: hnRNP A1; 42kDa:We are unsure as to the identity of this extra band.

References for Anti-hnRNP A1 antibody [9H10] (ab5832)

This product has been referenced in:
  • Le PN  et al. TERRA, hnRNP A1, and DNA-PKcs Interactions at Human Telomeres. Front Oncol 3:91 (2013). WB, IP ; Human . Read more (PubMed: 23616949) »
  • Singh NN  et al. An intronic structure enabled by a long-distance interaction serves as a novel target for splicing correction in spinal muscular atrophy. Nucleic Acids Res 41:8144-65 (2013). WB . Read more (PubMed: 23861442) »

See all 11 Publications for this product

Product Wall

Application Western blot
Loading amount 15 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native) (4-12% Bis-Tris Gel, MES running buffer)
Sample Human Cell lysate - whole cell (Human Embryonic Kidney)
Specification Human Embryonic Kidney
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted Mar 18 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Sample Rat Cell (oligodendrocytes)
Specification oligodendrocytes
Permeabilization Yes - 0,1%Triton X100
Fixative Paraformaldehyde
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Abcam user community

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Submitted Jan 03 2014

I have not been able to find the Drosophila sequence to check homology of the immunogen for either of these antibodies (full-length human hnRNP-A1) with the Drosophila sequence. This allows us to assess the likelihood of cross-reaction of the antibody ...

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Application Western blot
Sample Mouse Cell lysate - whole cell (NSC34)
Loading amount 10 µg
Specification NSC34
Gel Running Conditions Reduced Denaturing (4-12% nupage gel)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 13 2010

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (GM10725 Lymphoblastoid cell line)
Specification GM10725 Lymphoblastoid cell line
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Apr 02 2007

Application Western blot
Sample Human Cell lysate - whole cell (cell type: IDH4)
Loading amount 5 µg
Specification cell type: IDH4
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
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Miss. Elizabeth Kovacs

Verified customer

Submitted Jun 06 2006

Thank you for your enquiry. I am sorry to hear that your customer has been having problems with ab5832. Please thank them for taking the time to fill out our questionnaire. I have read their questionnaire and I have a few comments. Your protocol is...

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