Overview

  • Product name

    Anti-SF3B3 antibody [EPR18440]
    See all SF3B3 primary antibodies
  • Description

    Rabbit monoclonal [EPR18440] to SF3B3
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human SF3B3 aa 1150 to the C-terminus. The exact sequence is proprietary.
    Database link: Q15393

  • Positive control

    • WB: HeLa, Jurkat, C6 and RAW 264.7 whole cell lysates; Human fetal liver, fetal heart and fetal kidney lysates; Mouse brain, heart, kidney and spleen lysates; Rat brain, heart and kidney lysates. IHC-P: Human colon, Human breast cancer (ER+) and Human breast cancer (ER-) tissues; Mouse cerebral cortex tissue; Rat kidney tissue. ICC/IF: C6 and RAW 264.7 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab209402 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
IP 1/60.
WB 1/2000. Detects a band of approximately 136 kDa (predicted molecular weight: 136 kDa).
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.
Flow Cyt 1/200.

Target

  • Function

    Subunit of the splicing factor SF3B required for 'A' complex assembly formed by the stable binding of U2 snRNP to the branchpoint sequence (BPS) in pre-mRNA. Sequence independent binding of SF3A/SF3B complex upstream of the branch site is essential, it may anchor U2 snRNP to the pre-mRNA. May also be involved in the assembly of the 'E' complex. Belongs also to the minor U12-dependent spliceosome, which is involved in the splicing of rare class of nuclear pre-mRNA intron.
  • Sequence similarities

    Belongs to the RSE1 family.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • KIAA0017 antibody
    • Pre mRNA splicing factor SF3b 130 kDa subunit antibody
    • Pre-mRNA-splicing factor SF3b 130 kDa subunit antibody
    • RSE1 antibody
    • SAP 130 antibody
    • SAP130 antibody
    • SF3b130 antibody
    • SF3B3 antibody
    • SF3B3_HUMAN antibody
    • Spliceosome associated protein 130 antibody
    • Spliceosome-associated protein 130 antibody
    • Splicing factor 3b subunit 3 130kD antibody
    • Splicing factor 3B subunit 3 antibody
    • STAF130 antibody
    see all

Images

  • All lanes : Anti-SF3B3 antibody [EPR18440] (ab209402) at 1/20000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 136 kDa
    Observed band size: 136 kDa


    Exposure time: 8 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-SF3B3 antibody [EPR18440] (ab209402) at 1/2000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 136 kDa
    Observed band size: 136 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time : Lane 1/2: 4 seconds; Lane 3: 1 second.

  • All lanes : Anti-SF3B3 antibody [EPR18440] (ab209402) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat kidney lysate
    Lane 7 : Rat heart lysate
    Lane 8 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 9 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 136 kDa
    Observed band size: 136 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time : Lane 1/2/3: 4 seconds: Lane 4/5/6/8/9: 1 second; Lane 7: 8 seconds.

  • ab209402 at 1/60 immunoprecipitating SF3B3 in HeLa (human cervix adenocarcinoma) whole cell lysate observed at 136 KDa (lanes 1 and 2).

    Lane 1 (input): HeLa whole cell lysate 10μg

    Lane 2 (+): ab209402 + HeLa whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab209402 in HeLa whole cell lysate

    For western blotting, Panel A: ab209402, 1:500; Panel B: ab209403, 1:500 and VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Ab209402 was used to capture the target, and the WB was developed with ab209402 (A) ab209403 (B).

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

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling SF3B3 with ab209402 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus staining on Human colon 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.

  • Immunohistochemical analysis of paraffin-embedded Human breast cancer (ER+) tissue labeling SF3B3 with ab209402 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus staining on ER+ breast cancer 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.

  • Immunohistochemical analysis of paraffin-embedded Human breast cancer (ER-) tissue labeling SF3B3 with ab209402 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Weak nucleus staining on ER- breast cancer 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.

  • Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling SF3B3 with ab209402 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on mouse cerebral cortex 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.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling SF3B3 with ab209402 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nucleus staining on rat kidney 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.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cell line) cells labeling SF3B3 with ab209402 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on C6 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control  (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab209402 at 1/500 dilution followed by ab150120  at 1/1000 dilution.

    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling SF3B3 with ab209402 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on RAW 264.7 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG  (AlexaFluor®594) preadsorbed (ab150120) at  1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab209402 at 1/500 dilution followed by ab150120  at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling SF3B3 with ab209402 at 1/200 dilution (red) compared with a Rabbit IgG, monoclonal - Isotype control (ab172730)(black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

References

ab209402 has not yet been referenced specifically in any publications.

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