Overview

  • Product name

    Anti-CstF-64 antibody [EPR15698]
    See all CstF-64 primary antibodies
  • Description

    Rabbit monoclonal [EPR15698] to CstF-64
  • 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 CstF-64 aa 350-450. The exact sequence is proprietary.
    Database link: P33240

  • Positive control

    • WB: Jurkat, K562, THP-1 and HeLa whole cell lysates; Human testis tissue lysate. IHC: Human cervix carcinoma and Human tonsil tissue. ICC/IF: HeLa and Jurkat cells. IP: K562 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

  • 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

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 0.05% BSA, 40% Glycerol
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR15698
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab200837 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 1/500.
IP 1/30.
IHC-P 1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/5000. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).
Flow Cyt 1/20.

Target

  • Function

    One of the multiple factors required for polyadenylation and 3'-end cleavage of mammalian pre-mRNAs. This subunit is directly involved in the binding to pre-mRNAs.
  • Sequence similarities

    Contains 1 RRM (RNA recognition motif) domain.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization

    Nucleus. Localized with DDX1 in cleavage bodies.
  • Information by UniProt
  • Database links

  • Alternative names

    • betaCstF 64 variant 2 antibody
    • CF-1 64 kDa subunit antibody
    • CF1 64 kDa subunit antibody
    • Cleavage stimulation factor 2 antibody
    • Cleavage stimulation factor 64 kDa subunit antibody
    • Cleavage stimulation factor subunit 2 antibody
    • Cleavage stimulation factor, 3' pre RNA, subunit 2, 64kDa antibody
    • CSTF 2 antibody
    • CstF 64 antibody
    • CSTF 64 kDa subunit antibody
    • CstF-64 antibody
    • CSTF2 antibody
    • CSTF2_HUMAN antibody
    • MGC188397 antibody
    see all

Images

  • All lanes : Anti-CstF-64 antibody [EPR15698] (ab200837) at 1/5000 dilution

    Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 3 : THP-1 (Human monocytic leukemia cells) whole cell lysate
    Lane 4 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 61 kDa
    Observed band size: 61 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Flow Cytometry analysis of Jurkat (human acute T cell leukemia) labelling CstF-64 with purified ab200837 at 1/25000 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Alexa Fluor® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

     

  • Anti-CstF-64 antibody [EPR15698] (ab200837) at 1/200 dilution + Human testis tissue lysate

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

    Predicted band size: 61 kDa
    Observed band size: 61 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-CstF-64 antibody [EPR15698] (ab200837) at 1/5000 dilution

    Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate
    Lane 2 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with CSTF2 peptide
    Lane 3 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with CSTFT peptide

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 61 kDa
    Observed band size: 61 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Based on sequence analysis, ab200837 shares 78% homology with family member CSTFT. The levels of XR were tested in the accompanying peptide blocking experiment.

  • Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling CstF-64 with ab200837 at 1/100 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.

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

  • Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CstF-64 with ab200837 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on Human tonsil tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only: 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 HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling CstF-64 with ab200837 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear staining on HeLa cell line is observed. 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: ab200837 at 1/500 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.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling CstF-64 with ab200837 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear staining on Jurkat cell line is observed. 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: ab200837 at 1/500 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.

  • CstF-64 was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with ab200837 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab200837 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: K562 whole cell lysate10 µg (Input). Lane 2: ab200837 IP in K562 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200837 in K562 whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 5 seconds

References

ab200837 has not yet been referenced specifically in any publications.

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