Overview

  • Product name

    Anti-SF2 antibody [EPR8240]
    See all SF2 primary antibodies
  • Description

    Rabbit monoclonal [EPR8240] to SF2
  • Host species

    Rabbit
  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human SF2 aa 150-250 (internal sequence). The exact sequence is proprietary.

  • Positive control

    • IHC-P: Human colon and stomach tissue; WB: 293T, HeLa. K-562, RAW 264.7 and C6 cell lysates; ICC/IF: HeLa cells; FC:HeLa cells.
  • General notes

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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.
  • Dissociation constant (KD)

    KD = 5.70 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR8240
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab133689 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
WB 1/1000 - 1/10000. Predicted molecular weight: 27 kDa.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

ICC/IF 1/50 - 1/100.
Flow Cyt 1/40.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Plays a role in preventing exon skipping, ensuring the accuracy of splicing and regulating alternative splicing. Interacts with other spliceosomal components, via the RS domains, to form a bridge between the 5'- and 3'-splice site binding components, U1 snRNP and U2AF. Can stimulate binding of U1 snRNP to a 5'-splice site-containing pre-mRNA. Binds to purine-rich RNA sequences, either the octamer, 5'-RGAAGAAC-3' (r=A or G) or the decamers, AGGACAGAGC/AGGACGAAGC. Binds preferentially to the 5'-CGAGGCG-3' motif in vitro. Three copies of the octamer constitute a powerful splicing enhancer in vitro, the ASF/SF2 splicing enhancer (ASE) which can specifically activate ASE-dependent splicing. Isoform ASF-2 and isoform ASF-3 act as splicing repressors.
    • Sequence similarities

      Belongs to the splicing factor SR family.
      Contains 2 RRM (RNA recognition motif) domains.
    • Domain

      The RRM 2 domain plays an important role in governing both the binding mode and the phosphorylation mechanism of the RS domain by SRPK1. RS domain and RRM 2 are uniquely positioned to initiate a highly directional (C-terminus to N-terminus) phosphorylation reaction in which the RS domain slides through an extended electronegative channel separating the docking groove of SRPK1 and the active site. RRM 2 binds toward the periphery of the active site and guides the directional phosphorylation mechanism. Both the RS domain and an RRM domain are required for nucleocytoplasmic shuttling.
    • Post-translational
      modifications

      Phosphorylated by CLK1, CLK2, CLK3 and CLK4. Phosphorylated by SRPK1 at multiple serines in its RS domain via a directional (C-terminal to N-terminal) and a dual-track mechanism incorporating both processive phosphorylation (in which the kinase stays attached to the substrate after each round of phosphorylation) and distributive phosphorylation steps (in which the kinase and substrate dissociate after each phosphorylation event). The RS domain of SRSF1 binds to a docking groove in the large lobe of the kinase domain of SRPK1 and this induces certain structural changes in SRPK1 and/or RRM 2 domain of SRSF1, allowing RRM 2 to bind the kinase and initiate phosphorylation. The cycles continue for several phosphorylation steps in a processive manner (steps 1-8) until the last few phosphorylation steps (approximately steps 9-12). During that time, a mechanical stress induces the unfolding of the beta-4 motif in RRM 2, which then docks at the docking groove of SRPK1. This also signals RRM 2 to begin to dissociate, which facilitates SRSF1 dissociation after phosphorylation is completed.
      Arg-97 is dimethylated, probably to asymmetric dimethylarginine.
    • Cellular localization

      Cytoplasm. Nucleus speckle. In nuclear speckles. Shuttles between the nucleus and the cytoplasm.
    • Information by UniProt
    • Database links

    • Alternative names

      • Alternative splicing factor 1 antibody
      • Alternative-splicing factor 1 antibody
      • arginine/serine-rich 1 antibody
      • ASF 1 antibody
      • ASF antibody
      • ASF-1 antibody
      • ASF1 antibody
      • FLJ53078 antibody
      • MGC5228 antibody
      • P33 subunit antibody
      • Pre mRNA splicing factor SF2 P33 subunit antibody
      • pre-mRNA-splicing factor SF2 antibody
      • Serine/arginine-rich splicing factor 1 antibody
      • SF2 antibody
      • SF2P33 antibody
      • SFRS1 antibody
      • Splicing factor 2 alternate splicing factor antibody
      • Splicing factor 2 antibody
      • Splicing factor antibody
      • Splicing factor arginine/serine rich 1 antibody
      • SR Splicing factor 1 antibody
      • SRp30a antibody
      • srsf1 antibody
      • SRSF1_HUMAN antibody
      see all

    Images

    • All lanes : Anti-SF2 antibody [EPR8240] (ab133689) at 1/1000 dilution (Purified)

      Lane 1 : 293T (Human embryonic kidney epithelial cell) whole cell lysates
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 3 : C6 (Rat glial tumor glial cell) whole cell lysates
      Lane 4 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
      Lane 5 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 27 kDa
      Observed band size: 27 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue sections labeling SF2 with Purified ab133689 at 1/100 dilution (4.16 µg/ml). Heat mediated antigen retrieval was performed perform heat mediated antigen retrieval using Citrate buffer, pH 6.0. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

    • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling SF2 with purified ab133689 at 1/50 dilution (8.3 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling SF2 with purified ab133689 at 1/40 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    • All lanes : Anti-SF2 antibody [EPR8240] (ab133689) at 1/1000 dilution (unpurified)

      Lane 1 : 293T cell lysate
      Lane 2 : C6 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 27 kDa

    • Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling SF2 with unpurified ab133689 at 1/50 dilution.

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:

    • da Glória VG  et al. T cell activation regulates CD6 alternative splicing by transcription dynamics and SRSF1. J Immunol 193:391-9 (2014). Read more (PubMed: 24890719) »
    See 1 Publication for this product

    Customer reviews and Q&As

    Application
    Western blot
    Sample
    Zebrafish Tissue lysate - whole (embryonic tissue)
    Gel Running Conditions
    Reduced Denaturing (4-12% NuPAGE)
    Loading amount
    40000 cells
    Treatment
    50pg overexpressed flag-tagged construct
    Specification
    embryonic tissue
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Oct 21 2015

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