Overview

  • Product name

    Anti-SC35 antibody [EPR12238]
    See all SC35 primary antibodies
  • Description

    Rabbit monoclonal [EPR12238] to SC35
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human SC35 aa 50-150. The exact sequence is proprietary.
    Database link: Q01130

  • Positive control

    • WB: K562, HeLa, 293 and HepG2 cell lysates; Human thymus, fetal kidney and fetal brain lysates. IHC-P: Human cervical carcinoma tissue. ICC/IF: HEK-293 and HeLa cells. IP: HeLa whole cell lysate.
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab204916 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.
IP 1/30.
IHC-P 1/1000. 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: 25 kDa).
Flow Cyt 1/400.

Target

  • Function

    Necessary for the splicing of pre-mRNA. It is required for formation of the earliest ATP-dependent splicing complex and interacts with spliceosomal components bound to both the 5'- and 3'-splice sites during spliceosome assembly. It also is required for ATP-dependent interactions of both U1 and U2 snRNPs with pre-mRNA. 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. Binds to purine-rich RNA sequences, either 5'-AGSAGAGTA-3' (S=C or G) or 5'-GTTCGAGTA-3'. Can bind to beta-globin mRNA and commit it to the splicing pathway.
  • Sequence similarities

    Belongs to the splicing factor SR family.
    Contains 1 RRM (RNA recognition motif) domain.
  • Post-translational
    modifications

    Extensively phosphorylated on serine residues in the RS domain.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • 35 kDa antibody
    • arginine/serine-rich 2 antibody
    • PR264 antibody
    • Protein PR264 antibody
    • SC 35 antibody
    • SC-35 antibody
    • SC35 antibody
    • Serine/arginine-rich splicing factor 2 antibody
    • SFRS 2 antibody
    • SFRS2 antibody
    • SFRS2A antibody
    • Splicing component 35 kDa antibody
    • Splicing component antibody
    • Splicing factor antibody
    • Splicing factor arginine/serine rich 2 antibody
    • Splicing factor SC35 antibody
    • Splicing speckle antibody
    • Splicing speckles antibody
    • SR splicing factor 2 antibody
    • SRp30b antibody
    • SRSF2 antibody
    • SRSF2_HUMAN antibody
    see all

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293 (Human epithelial cells from embryonic kidney) cells labeling SC35 with ab204916 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HEK-293 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab204916 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • All lanes : Anti-SC35 antibody [EPR12238] (ab204916) at 1/1000 dilution

    Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 3 : 293 (Human epithelial cells from embryonic kidney) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 25 kDa
    Observed band size: 35 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labeling SC35 with ab204916 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human cervical carcinoma is observed. Counter stained with Hematoxylin.

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

  • SC35 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab204916 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab204916 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.

    Lane 1: HeLa whole cell lysate 10ug (Input). Lane 2: ab204916 IP in HeLa whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab204916 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

  • Anti-SC35 antibody [EPR12238] (ab204916) at 1/1000 dilution + HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 20 µg

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

    Predicted band size: 25 kDa
    Observed band size: 35 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling SC35 with ab204916 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab204916 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Anti-SC35 antibody [EPR12238] (ab204916) at 1/5000 dilution + Human thymus lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 25 kDa
    Observed band size: 35 kDa why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-SC35 antibody [EPR12238] (ab204916) at 1/1000 dilution + Human fetal kidney lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 25 kDa
    Observed band size: 35 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-SC35 antibody [EPR12238] (ab204916) at 1/1000 dilution + Human fetal brain lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 25 kDa
    Observed band size: 35 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

References

This product has been referenced in:

  • Teslow EA  et al. Obesity-induced MBD2_v2 expression promotes tumor-initiating triple-negative breast cancer stem cells. Mol Oncol 13:894-908 (2019). Read more (PubMed: 30636104) »
  • Chakraborty P  et al. DHX9 helicase promotes R-loop formation in cells with impaired RNA splicing. Nat Commun 9:4346 (2018). Read more (PubMed: 30341290) »
See all 3 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Zebrafish Cell (Whole Embryo, Dome Stage)
Permeabilization
Yes - 100% Methanol -20°C overnight, ddH2O room temperature for 15 minutes
Specification
Whole Embryo, Dome Stage
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Formaldehyde

Dr. Lennart Hilbert

Verified customer

Submitted Sep 23 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (Human lymphocytes)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
40 µg
Specification
Human lymphocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 27 2016

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