Recombinant
RabMAb

Recombinant Anti-SF2 antibody [EPR8239] (Alexa Fluor® 647) (ab197878)

Overview

  • Product name

    Anti-SF2 antibody [EPR8239] (Alexa Fluor® 647)
    See all SF2 primary antibodies
  • Description

    Rabbit monoclonal [EPR8239] to SF2 (Alexa Fluor® 647)
  • Host species

    Rabbit
  • Conjugation

    Alexa Fluor® 647. Ex: 652nm, Em: 668nm
  • Tested applications

    Suitable for: Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

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

  • Positive control

    • ICC/IF: HepG2 cells. Flow Cyt: HepG2 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.

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

    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. Store In the Dark.
  • Dissociation constant (KD)

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

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: 30% Glycerol, 1% BSA, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR8239
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab197878 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 1/50.

ab199093 - Rabbit monoclonal IgG (Alexa Fluor® 647), is suitable for use as an isotype control with this antibody.

ICC/IF 1/100.

This product gave a positive signal in HepG2 cells fixed with 4% formaldehyde (10 min)

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

  • Overlay histogram showing HepG2 cells stained with ab197878 (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 (ab197878, 1/50 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 647 used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a solid-state 25mW red diode laser (635 nm) and 675/30 bandpass filter.

  • ab197878 staining SF2 in HepG2 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab197878 at a 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at a 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

References

ab197878 has not yet been referenced specifically in any publications.

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