Overview

  • Product name

    Anti-SF2 antibody [EPR8239]
    See all SF2 primary antibodies
  • Description

    Rabbit monoclonal [EPR8239] to SF2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human SF2 aa 1-100. The exact sequence is proprietary.

  • Positive control

    • WB: HepG2, Raji, HeLa, 293T cell lysates; mouse spleen. IHC-P: Human prostatic hyperplasia and breast carcinoma, rat kidney and mouse cerebral cortex tissues. ICC/IF: HeLa cells. Flow Cyt: 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. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 8.00 x 10 -12 M
    Learn more about KD
  • Storage buffer

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

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR8239
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab129108 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 at an assay dependent concentration.
WB 1/10000. Detects a band of approximately 22-32 kDa (predicted molecular weight: 28 kDa).
IHC-P 1/800 - 1/1500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.

ICC/IF 1/150 - 1/500.

 

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

  • Anti-SF2 antibody [EPR8239] (ab129108) at 1/10000 dilution (purified) + Mouse spleen tissue at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 28 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic hyperplasia tissue labelling SF2 with purified ab129108 at 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Flow Cytometry analysis of HeLa cells labelling SF2 with purified ab129108 at a dilution of 1/70 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Flour® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling SF2 with purified ab129108 at 1/150. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Counterstained with DAPI.

    Control - primary antibody (1/150), secondary antibody ab150120 an Alexa Fluor® 594-conjugate goat anti-mouse IgG (1/500).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling SF2 with purified ab129108 at 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebral cortex tissue labelling SF2 with purified ab129108 at 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • All lanes : Anti-SF2 antibody [EPR8239] (ab129108) at 1/10000 dilution (purified)

    Lane 1 : HepG2 cell lysate
    Lane 2 : Raji cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : 293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 28 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling SF2 with unpurified ab129108 at 1/100.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:

  • Zhai W  et al. Sunitinib-suppressed miR-452-5p facilitates renal cancer cell invasion and metastasis through modulating SMAD4/SMAD7 signals. Mol Cancer 17:157 (2018). Read more (PubMed: 30419914) »
  • García-Berrocoso T  et al. Single Cell Immuno-Laser Microdissection Coupled to Label-Free Proteomics to Reveal the Proteotypes of Human Brain Cells After Ischemia. Mol Cell Proteomics 17:175-189 (2018). Read more (PubMed: 29133510) »
See all 9 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

1-2 of 2 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U-2OS)
Permeabilization
No
Specification
U-2OS
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Methanol

Abcam user community

Verified customer

Submitted Jun 13 2017

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Dermal Fibroblast)
Total protein in input
600 µg
Immuno-precipitation step
Protein G
Specification
Dermal Fibroblast

Abcam user community

Verified customer

Submitted Jul 19 2016

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up