• Product name
  • Description
    Rabbit polyclonal to U2AF65
  • Host species
  • Tested applications
    Suitable for: IHC-P, ChIP, ICC/IF, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human, Zebrafish
    Predicted to work with: Cow, Xenopus laevis
  • Immunogen

    Synthetic peptide derived from within residues 200 - 300 of Human U2AF65.

    Read Abcam's proprietary immunogen policy (Peptide available as ab37529.)

  • Positive control
    • This antibody gave a positive result in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) A431 (Human epithelial carcinoma cell line) HEK 293 (Human embryonic kidney cell line) HepG2 (Human hepatocellular liver carcinoma cell line) MCF-7 (Human breast adenocarcinoma cell line) SHSY-5Y (Human neuroblastoma cell line) U2OS (Human osteosarcoma cell line)


  • Form
  • 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: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
  • Isotype
  • Research areas


Our Abpromise guarantee covers the use of ab37530 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
IHC-P Use a concentration of 5 µg/ml.
ChIP Use at an assay dependent concentration. PubMed: 27252533
ICC/IF Use a concentration of 1 µg/ml.
IP Use at an assay dependent concentration. PubMed: 21725067
WB 1/250. Detects a band of approximately 65 kDa (predicted molecular weight: 53 kDa).


  • Function
    Necessary for the splicing of pre-mRNA. Induces cardiac troponin-T (TNNT2) pre-mRNA exon inclusion in muscle. Regulates the TNNT2 exon 5 inclusion through competition with MBNL1. Binds preferentially to a single-stranded structure within the polypyrimidine tract of TNNT2 intron 4 during spliceosome assembly. Required for the export of mRNA out of the nucleus, even if the mRNA is encoded by an intron-less gene. Represses the splicing of MAPT/Tau exon 10.
  • Sequence similarities
    Belongs to the splicing factor SR family.
    Contains 3 RRM (RNA recognition motif) domains.
  • Post-translational
    Lysyl-hydroxylation at Lys-15 and Lys-276 affects the mRNA splicing activity of the protein, leading to regulate some, but not all, alternative splicing events.
  • Cellular localization
  • Information by UniProt
  • Database links
  • Alternative names
    • hU2AF(65) antibody
    • hU2AF65 antibody
    • Splicing factor U2AF 65 kDa subunit antibody
    • U2 (RNU2) small nuclear RNA auxiliary factor 2 antibody
    • U2 auxiliary factor 65 kDa subunit antibody
    • U2 small nuclear ribonucleoprotein auxiliary factor (65kD) antibody
    • U2 small nuclear RNA auxiliary factor 2 antibody
    • U2 snRNP auxiliary factor large subunit antibody
    • U2af2 antibody
    • U2AF2_HUMAN antibody
    • U2AF65 antibody
    see all


  • All lanes : Anti-U2AF65 antibody (ab37530) at 1/250 dilution

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : A431 whole cell lysate (ab7909)
    Lane 4 : HEK293 whole cell lysate (ab7902)
    Lane 5 : HepG2 whole cell lysate (ab7900)
    Lane 6 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
    Lane 8 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    All lanes : Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 53 kDa
    Observed band size: 65 kDa
    why is the actual band size different from the predicted?

    Although the predicted band size is 53kDa based on Swiss-prot data, a band of 65kDa has been previously observed.  J Biol Chem. 2004 Nov 26;279(48):49773-9 (PMID: 15377657)

  • ICC/IF image of ab37530 stained human HeLa cells. The cells were PFA fixed (10 min) and incubated with the antibody (ab37530, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used to quench autofluorescence.  5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • IHC image of ab37530 staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab37530, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • All lanes : Anti-U2AF65 antibody (ab37530) at 1/250 dilution

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate at 20 µg
    Lane 3 : Zebrafish heart homogenate at 10 µg
    Lane 4 : Zebrafish liver homogenate at 10 µg
    Lane 5 : Zebrafish skeletal muscle homogenate at 10 µg
    Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

    All lanes : Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 53 kDa
    Observed band size: 53 kDa

    Exposure time: 5 minutes
  • U2AF65 was immunoprecipitated using 0.5mg SHSY5Y whole cell extract, 5µg of Rabbit polyclonal to U2AF65 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, SHSY5Y 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 ab37530.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 65kDa: U2AF65.


This product has been referenced in:
  • Yuan T  et al. Low-density lipoprotein receptor-related protein 6 regulates alternative pre-mRNA splicing. J Cell Mol Med 22:4653-4663 (2018). Read more (PubMed: 30070011) »
  • Rodríguez-Mateo C  et al. Downregulation of Lnc-Spry1 mediates TGF-ß-induced epithelial-mesenchymal transition by transcriptional and posttranscriptional regulatory mechanisms. Cell Death Differ 24:785-797 (2017). Read more (PubMed: 28186499) »
See all 12 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Western blot
Human Cell lysate - whole cell (Human Fibroblast)
Loading amount
20 µg
Human Fibroblast
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris Nupage gel)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 09 2013

Immunocytochemistry/ Immunofluorescence
Chinese Hamster Cell (CHO cells)
CHO cells
Yes - ETOH
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: RT°C

Mrs. Diana Vargas

Verified customer

Submitted Feb 10 2011


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