Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Caspase-7 antibody [E22] (ab32522)

Key features and details

  • Rabbit monoclonal [E22] to Caspase-7
  • Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IP
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-Caspase-7 antibody [E22]
    See all Caspase-7 primary antibodies

  • Description

    Rabbit monoclonal [E22] to Caspase-7

  • Host species

    Rabbit

  • Specificity

    The antibody should recognize both pro-form and p20 cleaved-form. The antibody does not cross-react with other Caspase family members.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: Jurkat and HeLa whole cell lysate (ab150035). IHC-P: Human skin cancer tissue. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: Jurkat cell lysates

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    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

Images

  • Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
    Western blot - Anti-Caspase-7 antibody [E22] (ab32522)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: Wild type HAP1 + Staurosporine ab120056 whole cell lysate (20 µg)
    Lane 3: CASP7 knockout HAP1 whole cell lysate (20 µg)
    Lane 4: CASP7 + Staurosporine knockout HAP1 whole cell lysate (20 µg)
    Lane 5: HeLa whole cell lysate (20 µg)
    Lane 6: HeLa + Staurosporine whole cell lysate (20 µg)

    Lanes 1 - 6: Merged signal (red and green). Green - ab32522 observed at 38 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab32522 was shown to specifically react with HAP1 + Staurosporine  when HAP1 + Staurosporine knockout samples were used. Wild-type and HAP1 + Staurosporine knockout samples were subjected to SDS-PAGE. Ab32522 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [E22] (ab32522)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [E22] (ab32522)

    Immunohistochemical analysis of paraffin embedded human skin cancer tissue using ab32522 at 1:50 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-Caspase-7 antibody [E22] (ab32522)
    Immunoprecipitation - Anti-Caspase-7 antibody [E22] (ab32522)

    Purified ab32522 at 1/20 dilution (1µg) immunoprecipitating Caspase-7 in Jurkat whole cell lysate.
    Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10µg
    Lane 2 (+): ab32522 + Jurkat whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32522 in Jurkat whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 34 kDa

  • Flow Cytometry - Anti-Caspase-7 antibody [E22] (ab32522)
    Flow Cytometry - Anti-Caspase-7 antibody [E22] (ab32522)

    Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Caspase-7 (red) with ab32522 at a 1/250 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

  • Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
    Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
    All lanes : Anti-Caspase-7 antibody [E22] (ab32522) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : CASP7 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 34 kDa
    Observed band size: 38 kDa
    why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab32522 observed at 38 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab32522 was shown to react with pro Caspase-7 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265777 (knockout cell lysate ab257380) was used. Wild-type HeLa and CASP7 knockout HeLa cell lysates were subjected to SDS-PAGE. ab32522 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
    Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
    Anti-Caspase-7 antibody [E22] (ab32522) at 1/1000 dilution + Jurkat cell lysate

    Predicted band size: 34 kDa
    Observed band size: 34 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Caspase-7 antibody [E22] (ab32522)
    Immunocytochemistry/ Immunofluorescence - Anti-Caspase-7 antibody [E22] (ab32522)

    Immunofluorescent staining of HeLa cells using ab32522 at 1:100 dilution.

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