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

Reviews (5)Q&A (2)References (39)
Western blot - Anti-hnRNP A1 antibody [9H10] (ab5832)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP A1 antibody [9H10] (ab5832)
  • Western blot - Anti-hnRNP A1 antibody [9H10] (ab5832)
  • Flow Cytometry - Anti-hnRNP A1 antibody [9H10] (ab5832)
  • Immunoprecipitation - Anti-hnRNP A1 antibody [9H10] (ab5832)

Key features and details

  • Mouse monoclonal [9H10] to hnRNP A1
  • Suitable for: Flow Cyt, IHC-P, WB, IP
  • Reacts with: Human
  • Isotype: IgG2b

Overview

  • Product name

    Anti-hnRNP A1 antibody [9H10]
    See all hnRNP A1 primary antibodies

  • Description

    Mouse monoclonal [9H10] to hnRNP A1

  • Host species

    Mouse

  • Tested applications

    Suitable for: Flow Cyt, IHC-P, WB, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • General notes

    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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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
    Constituent: 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

The Abpromise guarantee

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.
WB (3)
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.
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.
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.

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

    • Western blot - Anti-hnRNP A1 antibody [9H10] (ab5832)
      Western blot - Anti-hnRNP A1 antibody [9H10] (ab5832)
      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
      All lanes : 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
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP A1 antibody [9H10] (ab5832)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP A1 antibody [9H10] (ab5832)
      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.
    • Western blot - Anti-hnRNP A1 antibody [9H10] (ab5832)
      Western blot - Anti-hnRNP A1 antibody [9H10] (ab5832)This image is courtesy of an anonymous Abreview.
      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
      All lanes : IRDye® 700DX-conjugated Donkey anti-Mouse IgG at 1/3000 dilution

      Performed under reducing conditions.

      Predicted band size: 38 kDa


      Exposure time: 1 minute


      Knockdown for 72 hours.

      See Abreview

    • Flow Cytometry - Anti-hnRNP A1 antibody [9H10] (ab5832)
      Flow Cytometry - Anti-hnRNP A1 antibody [9H10] (ab5832)
      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.

    • Immunoprecipitation - Anti-hnRNP A1 antibody [9H10] (ab5832)
      Immunoprecipitation - Anti-hnRNP A1 antibody [9H10] (ab5832)
      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 (39)

    Publishing research using ab5832? Please let us know so that we can cite the reference in this datasheet.

    ab5832 has been referenced in 39 publications.

    • Möller K  et al. Upregulation of the heterogeneous nuclear ribonucleoprotein hnRNPA1 is an independent predictor of early biochemical recurrence in TMPRSS2:ERG fusion-negative prostate cancers. Virchows Arch N/A:N/A (2020). PubMed: 32417965
    • Sun G  et al. HnRNP A1 - mediated alternative splicing of CCDC50 contributes to cancer progression of clear cell renal cell carcinoma via ZNF395. J Exp Clin Cancer Res 39:116 (2020). PubMed: 32560659
    • Guo YE  et al. Pol II phosphorylation regulates a switch between transcriptional and splicing condensates. Nature 572:543-548 (2019). PubMed: 31391587
    • Tsai PL  et al. Methodologies to monitor protein turnover at the inner nuclear membrane. Methods Enzymol 619:47-69 (2019). PubMed: 30910029
    • Thompson MG  et al. Co-regulatory activity of hnRNP K and NS1-BP in influenza and human mRNA splicing. Nat Commun 9:2407 (2018). PubMed: 29921878
    View all Publications for this product

    Customer reviews and Q&As

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    1-7 of 7 Abreviews or Q&A

    Western blot abreview for Anti-hnRNP A1 antibody [9H10]

     Excellent
    Abreviews
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 18 2014

    Immunocytochemistry/ Immunofluorescence abreview for Anti-hnRNP A1 antibody [9H10]

     Inconclusive
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 03 2014

    Western blot abreview for Anti-hnRNP A1 antibody [9H10]

     Good
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 13 2010

    Immunocytochemistry/ Immunofluorescence abreview for Anti-hnRNP A1 antibody [9H10]

     Good
    Abreviews
    abreview image
    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%
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 02 2007

    Western blot abreview for Anti-hnRNP A1 antibody [9H10]

     Excellent
    Abreviews
    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%
    Read More

    Miss. Elizabeth Kovacs

    Verified customer

    Submitted Jun 06 2006

    Question

    Dear Tech Support Team, Our customer is willing to test ab5832 or ab10685 in IP with Drosophila samples. Are you interested in this type of the information for the Abreview promotion (The customer buys certain antibodies and upload Abreview on Abcam's website and then we give 100% credit note for this antibody to buy next antibody)? Do you have information about the homology of the immunogens of these 2 antibodies with Drosophila protein? Which of these two is preferable for the test withthe abreview promotion? Thanks in advance for your assistance and reply.

    Read More
    Answer

    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 with the untested species. I did find an apparent analog, the product of the Dm gene Hrb87F (Synonym: hrp36). Alignment of human hnRNP1 with this protein returns 57% identity. This suggests low potential for cross-reactivity (we look for 85% or better), but if your customer is still interested in trying one of these to IP Dm protein, I suggest ab5832, as there have been inconsistent results lately from ab10685. However, the antibody does not IP the hnRNP complex.

    Read More

    Question

    BATCH NUMBER 132644 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM no band detectable even after an exposure time of 20 min SAMPLE protein extract derived from peripheral blood mononuclear cells (PBMCs) PRIMARY ANTIBODY protein transfer to nitrocellulose membrane by wet blotting (over-night, 30V), standard transfer buffer based on methanol, after transfer: quality control by Ponceau-staining, blocking of membrane with 6% milk/TBS-Tween (based on non-fat dry milk) for 4 hours @ RT DETECTION METHOD SuperSignal West Pico Chemiluminescent Substrate (Pierce), detection with Hyperfilm ECL (Amersham) POSITIVE AND NEGATIVE CONTROLS USED the same membrane was stained with antibodies against beta-actin and SF2/ASF which worked great as usual; furthermore, the same protein extracts were used to stain SMN; hnRNP A1 is ubiquitously expressed and should therefore also be present in PBMCs; a positive control was not delivered with the antibody ANTIBODY STORAGE CONDITIONS as indicated SAMPLE PREPARATION isolation of PBMCs using BD Vacutainer? CPT Cell Preparation tubes, 2x washing of cells with 1xPBS, protein extraction with RIPA w/o protease inhibitors, sample storage @-80?C; volume corresponding to 7.5?g of protein/sample plus 2xLaemmli buffer, heating @95?C for 5 min, 12%SDS PAGE @50-100V for about 2.5h TRANSFER AND BLOCKING CONDITIONS protein transfer to nitrocellulose membrane by wet blotting (over-night, 30V), standard transfer buffer based on methanol, after transfer: quality control by Ponceau-staining, blocking of membrane with 6% milk/TBS-Tween (based on non-fat dry milk) for 4 hours @ RT SECONDARY ANTIBODY Peroxidase-conjugated Affiniti Pure Goat Anti-Mouse IgG (H+L)(competitor) 1:10.000 in 1.5% milk/TBS Tween (based on non-fat dry milk), @4?C, for 1h Washing: 5x5 min with TBS Tween HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No WHAT STEPS HAVE YOU ALTERED? An opportunity for alteration would be the use of higher amounts of the primary antibody, however, the total volume delivered is as low as 50 ?l. In our experimental set up, we used a 1:1000 dilution (10?l of antibody in 10ml total incubation volume). Switching to a 1:200 or 1:100 dilution is not possible with an antibody volume of 50?l delivered in total. Thus, alterations are only theoretically possible. ADDITIONAL NOTES the lab and me as investigator are experienced in the field of western blotting. no bands at all even after an exposure time of 20 min (besides of medium background which is present all over the blot) We would appreciate being reimbursed by another antibody aliquot of the same or another clone to hnRNP A1 which is working properly or by not paying for the aliquot we already got. A picture showing the result after 20 min exposure is available on request. Thank you very much for your efforts and your understanding!

    Read More
    Answer

    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 not dissimilar to one that I would recommend for this antibody. I would also recommend a RIPA extraction given hnRNP A1 is a nuclear protein. We have had a favourable review posted recently for this antiserum by Naoko Tanese. His review did not appear to indicate anything that was performed differently from the standard conditions. However, given your protocol please may I suggest that you repeat this experiment, extracting your cells once again, just to rule out the fact that hnRNP A1 was not extracted by the RIPA buffer. It would also be of benefit to prolong the incubation time to overnight at 4oC and increasing the total mass of protein loaded onto the gel to 20ug. Also if possible change from Milk as a blocker to 5% BSA as this can give cleaner results. I think that keeping the dilution at 1:1000 is Ok as again Naoko Tanese, in his review obtained good results with this dilution. I think that combining these conditions would serve to give you the best chance of obtaining some favourable results. I look forward to hearing from you.

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