Overview

  • Product name

    Anti-SF3B3 antibody [EPR18441]
    See all SF3B3 primary antibodies
  • Description

    Rabbit monoclonal [EPR18441] to SF3B3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human SF3B3 aa 1050-1150. The exact sequence is proprietary.
    Database link: Q15393

  • Positive control

    • WB: HeLa, Jurkat, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Human fetal liver and fetal heart lysates; Mouse brain, kidney and spleen lysates; Rat kidney and spleen 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. IP: HeLa and C6 cell lysates.
  • 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 ab209403 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
WB 1/2000. Detects a band of approximately 136 kDa (predicted molecular weight: 136 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.
IP 1/60.

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 [EPR18441] (ab209403) at 1/5000 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.

  • Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling SF3B3 with ab209403 at 1/250 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 ab209403 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: ab209403 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.

  • SF3B3 was immunoprecipitated from 1mg / 300µl of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab209403 at 1/60 dilution (5µg/ 1mg of lysate).

    Western blot was performed from the immunoprecipitate using ab209403 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate, 10ug (Input).

    Lane 2: ab209403 IP in HeLa whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal - Isotype Control (ab172730) instead of ab209403 in HeLa whole cell lysate.

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

    Exposure time: 1 second.

  • All lanes : Anti-SF3B3 antibody [EPR18441] (ab209403) at 1/2000 dilution

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


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-SF3B3 antibody [EPR18441] (ab209403) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat kidney lysate
    Lane 5 : Rat spleen lysate
    Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 7 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 8 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 9 : NIH/3T3 (Mouse embryonic fibroblast cell line) 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 1/2/4: 3 minutes; Lane 3/5: 30 seconds; Lane 6/7/8/9: 8 seconds.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling SF3B3 with ab209403 at 1/250 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.

  • Immunohistochemical analysis of paraffin-embedded Human breast cancer (ER+) tissue labeling SF3B3 with ab209403 at 1/250 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.

  • Immunohistochemical analysis of paraffin-embedded Human breast cancer (ER-) tissue labeling SF3B3 with ab209403 at 1/250 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 ab209403 at 1/250 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.

  • 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 ab209403 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: ab209403 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.

  • SF3B3 was immunoprecipitated from 1mg / 300µl of C6 (Rat glial tumor cell line) whole cell lysate with ab209403 at 1/60 dilution (5µg / 1mg of lysate).

    Western blot was performed from the immunoprecipitate using ab209403 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: C6 whole cell lysate, 10ug (Input).

    Lane 2: ab209403 IP in C6 whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal - Isotype Control (ab172730) instead of ab209403 in HeLa whole cell lysate.

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

    Exposure time: 1 second.

References

ab209403 has not yet been referenced specifically in any publications.

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