Recombinant
RabMAb

Recombinant Anti-PRPF4 antibody [EPR17206(B)] (ab201684)

Overview

  • Product name

    Anti-PRPF4 antibody [EPR17206(B)]
    See all PRPF4 primary antibodies
  • Description

    Rabbit monoclonal [EPR17206(B)] to PRPF4
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human PRPF4 aa 1-100. The exact sequence is proprietary.
    Database link: O43172

  • Positive control

    • WB: HeLa, SW480, Raji and Jurkat whole cell lysates; Human fetal spleen lysate. IHC-P: Human cervix carcinoma tissue. ICC/IF: HeLa and MCF7 cells. IP and FC: 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 ab201684 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 1/5000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
IHC-P 1/3000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1200.
IP 1/40.
Flow Cyt 1/50.

Target

  • Function

    Participates in pre-mRNA splicing. Part of the U4/U5/U6 tri-snRNP complex, one of the building blocks of the spliceosome.
  • Sequence similarities

    Contains 7 WD repeats.
  • Cellular localization

    Nucleus speckle. Colocalizes with spliceosomal snRNPs.
  • Information by UniProt
  • Database links

  • Alternative names

    • hPrp4 antibody
    • HPRP4P antibody
    • PRP4 antibody
    • PRP4 homolog antibody
    • PRP4 pre mRNA processing factor 4 homolog (yeast) antibody
    • PRP4/STK/WD splicing factor antibody
    • PRP4_HUMAN antibody
    • Prp4p antibody
    • PRPF4 antibody
    • RP70 antibody
    • SNRNP60 antibody
    • U4/U6 small nuclear ribonucleoprotein Prp4 antibody
    • U4/U6 snRNP 60 kDa protein antibody
    • WD splicing factor Prp4 antibody
    see all

Images

  • All lanes : Anti-PRPF4 antibody [EPR17206(B)] (ab201684) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : SW480 (Human colon adenocarcinoma cell line) whole cell lysate
    Lane 3 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
    Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) 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: 58 kDa
    Observed band size: 58 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling PRPF4 with ab201684 at 1/3000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on Human cervix carcinoma tissue 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.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PRPF4 with ab201684 at 1/1200 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 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 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab201684 at 1/1200 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 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 1/500 dilution.

     

  • Anti-PRPF4 antibody [EPR17206(B)] (ab201684) at 1/5000 dilution + Human fetal spleen lysate at 10 µg

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

    Predicted band size: 58 kDa
    Observed band size: 58 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling PRPF4 with ab201684 at 1/1200 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on MCF7 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 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab201684 at 1/1200 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 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 1/500 dilution.

     

  • Flow cytometry analysis of HeLa cells labelling PRPF4 (red) with purified ab201684 at dilution of 1/50. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody. 

  • PRPF4 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab201684 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab201684 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1500 dilution.

    Lane 1: HeLa whole cell lysate 10 µg (Input). Lane 2: ab201684 IP in HeLa whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab201684 in HeLa whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 5 seconds.

References

ab201684 has not yet been referenced specifically in any publications.

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