Recombinant
RabMAb

Recombinant Anti-SC35 antibody [EPR12238] (ab204916)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR12238] to SC35
  • Suitable for: ICC/IF, IP, IHC-P, WB, Flow Cyt
  • Reacts with: Human

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

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

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

    • Immunocytochemistry/ Immunofluorescence - Anti-SC35 antibody [EPR12238] (ab204916)
      Immunocytochemistry/ Immunofluorescence - Anti-SC35 antibody [EPR12238] (ab204916)

      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.

    • Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
      Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
      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.

    • Flow Cytometry - Anti-SC35 antibody [EPR12238] (ab204916)
      Flow Cytometry - Anti-SC35 antibody [EPR12238] (ab204916)

      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling SC35 with ab204916 at 1:400 dilution (0.1μg)/ Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1:2000 dilution was used as the secondary antibody.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SC35 antibody [EPR12238] (ab204916)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SC35 antibody [EPR12238] (ab204916)

      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.

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

    • Immunoprecipitation - Anti-SC35 antibody [EPR12238] (ab204916)
      Immunoprecipitation - Anti-SC35 antibody [EPR12238] (ab204916)

      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.

    • Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
      Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-SC35 antibody [EPR12238] (ab204916)
      Immunocytochemistry/ Immunofluorescence - Anti-SC35 antibody [EPR12238] (ab204916)

      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.

    • Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
      Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
      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.

    • Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
      Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
      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.

    • Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
      Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
      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

    Publishing research using ab204916? Please let us know so that we can cite the reference in this datasheet.

    ab204916 has been referenced in 5 publications.

    • Teslow EA  et al. Obesity-induced MBD2_v2 expression promotes tumor-initiating triple-negative breast cancer stem cells. Mol Oncol 13:894-908 (2019). PubMed: 30636104
    • Gudkova D  et al. HCF-2 inhibits cell proliferation and activates differentiation-gene expression programs. Nucleic Acids Res 47:5792-5808 (2019). PubMed: 31049581
    • Li X  et al. Downregulation of SRPK2 promotes cell cycle arrest though E2F1 in non-small cell lung cancer. Eur J Histochem 63:N/A (2019). PubMed: 31833327
    • Chakraborty P  et al. DHX9 helicase promotes R-loop formation in cells with impaired RNA splicing. Nat Commun 9:4346 (2018). PubMed: 30341290
    • Shakyawar DK  et al. C3G dynamically associates with nuclear speckles and regulates mRNA splicing. Mol Biol Cell 29:1111-1124 (2018). PubMed: 29496966

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-SC35 antibody [EPR12238]

     Good
    Abreviews
    abreview image
    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
    Read More

    Dr. Lennart Hilbert

    Verified customer

    Submitted Sep 23 2016

    Western blot abreview for Anti-SC35 antibody [EPR12238]

     Good
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 27 2016

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