Overview

  • Product name
    Anti-hnRNP A1 antibody [9H10]
    See all hnRNP A1 primary antibodies
  • Description
    Mouse monoclonal [9H10] to hnRNP A1
  • Tested applications
    Suitable for: Flow Cyt, IHC-P, ELISA, WB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

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

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.1% Sodium Azide
    Constituents: PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Purification notes
    Purified from tissue culture supernatant.
  • Clonality
    Monoclonal
  • Clone number
    9H10
  • Myeloma
    Sp2/0
  • Isotype
    IgG2b
  • 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 Abreviews Notes
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

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 concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 34 kDa (predicted molecular weight: 38 kDa).
IP Use at an assay dependent concentration. This antibody does not IP the hnRNP complex.
ICC/IF Use at an assay dependent concentration. See Abreview (April 2, 2007)

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

Anti-hnRNP A1 antibody [9H10] images

  • 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
  • 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 1/1000 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.

    Knockdown for 72 hours.

    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:
  • Meng X  et al. Heterogeneous nuclear ribonucleoprotein A1 interacts with microRNA-34a to promote chondrogenic differentiation of mesenchymal stem cells. Am J Transl Res 9:1774-1782 (2017). WB, RIP ; Mouse . Read more (PubMed: 28469782) »
  • Jiang P  et al. Fyn/heterogeneous nuclear ribonucleoprotein E1 signaling regulates pancreatic cancer metastasis by affecting the alternative splicing of integrin ß1. Int J Oncol 51:169-183 (2017). WB . Read more (PubMed: 28560430) »

See all 18 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
Username

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
Username

Abcam user community

Verified customer

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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Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam guarantees this product to work in the species/application used in this Abreview.
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%
Username

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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