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

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
  • Epigenetics and Nuclear Signaling
  • DNA / RNA
  • RNA Processing
  • Other

Applications

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
  modificationsArg-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
  • Clone ID: BC033714 Human
  • Entrez Gene: 3178 Human
  • Entrez Gene: 15382 Mouse
  • Omim: 164017 Human
  • SwissProt: P09651 Human
  • SwissProt: P49312 Mouse
  • Unigene: 546261 Human
  • Unigene: 655424 Human

  • Unigene: 237064 Mouse
  • Unigene: 432031 Mouse

  see all

• Alternative names
  HNRNPA 1 antibodyHelix destabilizing protein antibodyHelix-destabilizing protein antibody

  Heterogeneous nuclear ribonucleoprotein A1 antibodyHeterogeneous nuclear ribonucleoprotein A1B protein antibodyHeterogeneous nuclear ribonucleoprotein B2 protein antibodyHeterogeneous nuclear ribonucleoprotein core protein A1 antibodyhnRNP A1 antibodyhnRNP core protein A1 antibodyHNRNPA1 antibodyHNRPA1 antibodyMGC102835 antibodyNuclear ribonucleoprotein particle A1 protein antibodyROA1_HUMAN antibodySingle strand DNA binding protein UP1 antibodySingle strand RNA binding protein antibodySingle-strand RNA-binding protein antibodysyncrip antibody

  see all

Anti-hnRNP A1 antibody [9H10] images

• 

  Immunocytochemistry/ Immunofluorescence - hnRNP A1 antibody [9H10] (ab5832)This image is courtesy of an anonymous Abreview

  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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-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 - 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
  Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed (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
• 

  Western blot - hnRNP A1 antibody [9H10] (ab5832)
  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

• 

  Flow Cytometry-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)
  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 70^oC; 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.