Overview

  • Product name

    Anti-U2AF35/U2AF1 antibody [EPR12648(2)]
    See all U2AF35/U2AF1 primary antibodies
  • Description

    Rabbit monoclonal [EPR12648(2)] to U2AF35/U2AF1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human U2AF35/U2AF1 aa 50-150. The exact sequence is proprietary.
    Database link: Q01081

  • Positive control

    • WB: Ramos, HeLa and 293 whole cell lysates and rat spleen tissue lysate. IHC-P: Human transitional cell carcinoma of bladder and rat liver tissues. ICC/IF: Ramos cells. Flow Cyt: Ramos cells.
  • General notes

     This product was previously labelled as U2AF35

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR12648(2)
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab197591 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
ICC/IF 1/100.
Flow Cyt 1/170.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 35 kDa (predicted molecular weight: 28 kDa).

Target

  • Function

    Plays a critical role in both constitutive and enhancer-dependent splicing by mediating protein-protein interactions and protein-RNA interactions required for accurate 3'-splice site selection. Recruits U2 snRNP to the branch point. Directly mediates interactions between U2AF2 and proteins bound to the enhancers and thus may function as a bridge between U2AF2 and the enhancer complex to recruit it to the adjacent intron.
  • Sequence similarities

    Belongs to the splicing factor SR family.
    Contains 2 C3H1-type zinc fingers.
    Contains 1 RRM (RNA recognition motif) domain.
  • Domain

    The C-terminal SR-rich domain is required for interactions with SR proteins and the splicing regulators TRA and TRA2, and the N-terminal domain is required for formation of the U2AF1/U2AF2 heterodimer.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization

    Nucleus. Nucleus speckle.
  • Information by UniProt
  • Database links

  • Alternative names

    • DKFZp313J1712 antibody
    • FP793 antibody
    • Human U2 snRNP auxiliary factor small subunit9 antibody
    • RN antibody
    • RNU2AF1 antibody
    • Splicing factor U2AF 35 kDa subunit antibody
    • U2 auxiliary factor 35 kDa subunit antibody
    • U2 small nuclear ribonucleoprotein auxillary factor 35 KD subunit antibody
    • U2 small nuclear RNA auxiliary factor 1 antibody
    • U2 snRNP auxiliary factor small subunit antibody
    • U2(RNU2) small nuclear RNA auxiliary factor 1 antibody
    • U2(RNU2) small nuclear RNA auxiliary factor binding protein antibody
    • U2AF1 antibody
    • U2AF1_HUMAN antibody
    • U2AF35 antibody
    • U2AFBP antibody
    see all

Images

  • Anti-U2AF35/U2AF1 antibody [EPR12648(2)] (ab197591) at 1/10000 dilution + Ramos (Human Burkitt's lymphoma cell line) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 28 kDa
    Observed band size: 35 kDa
    why is the actual band size different from the predicted?



    Blocking/dilution buffer: 5% NFDM/TBST. 

    The observed MW is consistent with what has been described in the literature PMID:1388271.

  • All lanes : Anti-U2AF35/U2AF1 antibody [EPR12648(2)] (ab197591) at 1/20000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : 293 (Human embryonic kidney) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 28 kDa
    Observed band size: 35 kDa why is the actual band size different from the predicted?



    Blocking/dilution buffer: 5% NFDM/TBST. 

    The observed MW is consistent with what has been described in the literature PMID:1388271.

  • Anti-U2AF35/U2AF1 antibody [EPR12648(2)] (ab197591) at 1/1000 dilution + Rat spleen lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 28 kDa
    Observed band size: 35 kDa why is the actual band size different from the predicted?



    Blocking/dilution buffer: 5% NFDM/TBST.

    The observed MW is consistent with what has been described in the literature PMID:1388271

     

  • Immunohistochemical analysis of paraffin-embedded Human transitional cell carcinoma of bladder tissue labeling U2AF35/U2AF1 with ab197591 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm and nuclear staining on Human transitional cell carcinoma of bladder tissue is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling U2AF35/U2AF1 with ab197591 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP)(ab97051) secondary antibody at 1/500 dilution. Cytoplasm and nuclear staining on rat liver tissue is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/Immunofluorescence analysis of RAMOS cells labelling U2AF35/U2AF1 with ab197591 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
    Control: PBS only.
    Nuclear counter stain: DAPI.

  • Flow cytometric analysis of 2% paraformaldehyde-fixed Ramos (Human Burkitt's lymphoma) cells labeling U2AF35/U2AF1 with ab197591 at 1/170 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

References

ab197591 has not yet been referenced specifically in any publications.

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