Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-pro Caspase-3 antibody [E83-103] (ab32499)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E83-103] to pro Caspase-3
  • Suitable for: ICC/IF, WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-pro Caspase-3 antibody [E83-103]
    See all pro Caspase-3 primary antibodies

  • Description

    Rabbit monoclonal [E83-103] to pro Caspase-3

  • Host species

    Rabbit

  • Specificity

    This antibody only detects pro-form (35kD) of caspase-3, and does not recognize any cleaved caspases.

  • Tested Applications & Species

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

  • Immunogen

    Synthetic peptide corresponding to Human pro Caspase-3 (N terminal).

  • Positive control

    • Jurkat whole cell lysate (ab7899) and human colon adenocarcinoma.

  • General notes

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

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    E83-103

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-pro Caspase-3 antibody [E83-103] (ab32499)
    Western blot - Anti-pro Caspase-3 antibody [E83-103] (ab32499)

    Lane 1: Wild-type HAP1 cell lysate
    Lane 2: Wild-type HAP1 cell lysate + Staurosporine (1μM for 4h)
    Lane 3: Caspase-3 knockout HAP1 cell lysate
    Lane 4: Caspase-3 knockout HAP1 cell lysate + Staurosporine (1μM for 4h)
    Lanes 1 - 4: Merged signal (red and green). Green - ab32499 observed at 35 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab32499 was shown to specifically react with pro Caspase 3 when Caspase 3 knockout samples were used. Wild-type and Caspase 3 knockout samples (± Staurosporine treatment) were subjected to SDS-PAGE. ab32499 and ab8245 (loading control to GAPDH) were diluted to 1/1000 and 1/10000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pro Caspase-3 antibody [E83-103] (ab32499)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pro Caspase-3 antibody [E83-103] (ab32499)

    Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma ab32499 at 1/250 dilution.

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

  • Immunocytochemistry/ Immunofluorescence - Anti-pro Caspase-3 antibody [E83-103] (ab32499)
    Immunocytochemistry/ Immunofluorescence - Anti-pro Caspase-3 antibody [E83-103] (ab32499)

    ICC/IF image of ab32499 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32499, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, anti-rabbit DyLight® 488 used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Flow Cytometry - Anti-pro Caspase-3 antibody [E83-103] (ab32499)
    Flow Cytometry - Anti-pro Caspase-3 antibody [E83-103] (ab32499)

    Overlay histogram showing Jurkat cells stained with ab32499 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32499, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was anti-rabbit DyLight® 488 (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Western blot - Anti-pro Caspase-3 antibody [E83-103] (ab32499)
    Western blot - Anti-pro Caspase-3 antibody [E83-103] (ab32499)
    All lanes : Anti-pro Caspase-3 antibody [E83-103] (ab32499) at 1/10000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : Jurkat cell lysate + Camptothecin

    Predicted band size: 31 kDa
    Observed band size: 35 kDa
    why is the actual band size different from the predicted?

  • Anti-pro Caspase-3 antibody [E83-103] (ab32499)
    Anti-pro Caspase-3 antibody [E83-103] (ab32499)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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