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  1. Link

    sc35-antibody-epr12238-ab204916.pdf

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Epigenetics and Nuclear Signaling DNA / RNA RNA Processing Splicing
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RecombinantRabMAb

Recombinant Anti-SC35 antibody [EPR12238] (ab204916)

  • Datasheet
  • Certificate of Compliance
Reviews (2) Submit a question References (10)

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Immunocytochemistry/ Immunofluorescence - Anti-SC35 antibody [EPR12238] (ab204916)
  • Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
  • Flow Cytometry (Intracellular) - Anti-SC35 antibody [EPR12238] (ab204916)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SC35 antibody [EPR12238] (ab204916)
  • Immunoprecipitation - Anti-SC35 antibody [EPR12238] (ab204916)
  • Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
  • Immunocytochemistry/ Immunofluorescence - Anti-SC35 antibody [EPR12238] (ab204916)
  • Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
  • Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
  • Western blot - Anti-SC35 antibody [EPR12238] (ab204916)
  • Anti-SC35 antibody [EPR12238] (ab204916)

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: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WB
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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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: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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

  • 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.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR12238
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • Splicing
    • Tags & Cell Markers
    • Subcellular Markers
    • Nucleus
    • Other Nuclear Bodies

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-SC35 antibody [EPR12238] (ab222118)
    • Alexa Fluor® 647 Anti-SC35 antibody [EPR12238] (ab222919)
    • Anti-SC35 antibody [EPR12238] - BSA and Azide free (ab232656)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human SC35 protein (ab152674)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/400.
ICC/IF (1)
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)
1/1000. Detects a band of approximately 35 kDa (predicted molecular weight: 25 kDa).
Notes
Flow Cyt (Intra)
1/400.
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).

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.
  • Target information above from: UniProt accession Q01130 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6427 Human
    • Omim: 600813 Human
    • SwissProt: Q01130 Human
    • Unigene: 584801 Human
    • 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 (Intracellular) - Anti-SC35 antibody [EPR12238] (ab204916)
      Flow Cytometry (Intracellular) - Anti-SC35 antibody [EPR12238] (ab204916)

      Intracellular 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.

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

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (10)

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

    ab204916 has been referenced in 10 publications.

    • Tomita T  et al. Extracellular mRNA transported to the nucleus exerts translation-independent function. Nat Commun 12:3655 (2021). PubMed: 34135341
    • Yang L  et al. Mirtronic miR-4646-5p promotes gastric cancer metastasis by regulating ABHD16A and metabolite lysophosphatidylserines. Cell Death Differ 28:2708-2727 (2021). PubMed: 33875796
    • Chen Z  et al. CircPLCE1 facilitates the malignant progression of colorectal cancer by repressing the SRSF2-dependent PLCE1 pre-RNA splicing. J Cell Mol Med 25:7244-7256 (2021). PubMed: 34173324
    • Francis AC  et al. HIV-1 replication complexes accumulate in nuclear speckles and integrate into speckle-associated genomic domains. Nat Commun 11:3505 (2020). PubMed: 32665593
    • Ilik IA  et al. SON and SRRM2 are essential for nuclear speckle formation. Elife 9:N/A (2020). PubMed: 33095160
    • 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
    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
    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

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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