Overview

  • Product nameAnti-pro Caspase 3 antibody [E61]
    See all pro Caspase 3 primary antibodies
  • Description
    Rabbit monoclonal [E61] to pro Caspase 3
  • SpecificityThe antibody only recognizes the pro-form of Caspase-3. It does not react with the cleaved forms (active enzyme) of Caspase-3.
  • Tested applicationsSuitable for: Flow Cyt, IP, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human pro Caspase 3 (N terminal). The exact sequence is proprietary.

  • Positive control
    • HeLa cells lysate, cervical carcinoma tissue
  • General notes

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

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • Concentration information loading...
  • PurityIgG fraction
  • ClonalityMonoclonal
  • Clone numberE61
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab32150 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
Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/10.
WB 1/1000. Detects a band of approximately 35 kDa.
IHC-P Use at an assay dependent concentration.
ICC/IF 1/100.

Target

  • RelevanceCaspases are a family of cysteine proteases that are key mediators of programmed cell death or apoptosis. The precursor form of all caspases is composed of a prodomain, and large and small catalytic subunits. The active forms of caspases are generated by several stimuli including ligand-receptor interactions, growth factor deprivation and inhibitors of cellular functions. All known caspases require cleavage adjacent to aspartates to liberate one large and one small subunit, which associate into a2b2 tetramer to form the active enzyme. Gene for Caspase 3 also known as Yama, CPP32, and apopain codes for a 32-kDa protein. Caspase 3 cleaves the death substrate poly(ADP-ribose) polymerase (PARP) to a specific 85 kDa form observed during apoptosis and is inhibitable by the CrmA protein. Other Caspase 3 substrates include DNA-PK, actin, GAS2, and procaspase-6, etc. Caspase 3 is activated by cleavage events at Asp-28/Ser-29 (between N-terminal pro-domain) and Asp-175/Ser-176 (between large and small subunits) to generate a large subunit of 17-kDa and a small subunit of 12-kDa.
  • Cellular localizationCytoplasmic
  • Database links
  • Alternative names
    • CASP3 antibody
    • Caspase 3 antibody
    • Caspase 3 apoptosis related cysteine peptidase antibody
    • CPP32 antibody
    • CPP32B antibody
    • Procaspase 3 antibody
    • SCA 1 antibody
    • SCA1 antibody
    • Yama antibody
    • Yama protein antibody
    see all

Anti-pro Caspase 3 antibody [E61] images



  • Additional bands at : 35 kDa. We are unsure as to the identity of these extra bands.

    Lane 1: Wild-type HAP1 cell lysate
    Lane 2: Wild-type HAP1 cell lysate + Stuarosporine (1μM for 4h)
    Lane 3: Caspase-3 knockout HAP1 cell lysate
    Lane 4: Caspase-3 knockout HAP1 cell lysate + Stuarosporine (1μM for 4h)
    Lanes 1 - 4: Merged signal (red and green). Green - ab32150 observed at 35 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab32150 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. ab32150 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) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemical analysis of human paraffin-embedded cervical carcinoma tissue using ab32150 at 1/500 dilution.
  • Immunofluorescent staining of HeLa cells using ab32150 at 1/100 dilution.
  • Overlay histogram showing Jurkat cells stained with ab32150 (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. The cells were then incubated with the antibody (ab32150, 1/100 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 (1µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Anti-pro Caspase 3 antibody [E61] (ab32150) at 1/1000 dilution + Hela cell lysate

    Observed band size : 35 kDa (why is the actual band size different from the predicted?)

References for Anti-pro Caspase 3 antibody [E61] (ab32150)

This product has been referenced in:
  • Nilsonne G  et al. Phenotype-dependent apoptosis signalling in mesothelioma cells after selenite exposure. J Exp Clin Cancer Res 28:92 (2009). Flow Cyt ; Human . Read more (PubMed: 19563663) »

See 1 Publication for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (human C4-2 cells)
Loading amount 20 µg
Specification human C4-2 cells
Gel Running Conditions Reduced Denaturing
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Username

Mr. Hao Wu

Verified customer

Submitted Nov 21 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (293 FT cell line)
Loading amount 50 µg
Specification 293 FT cell line
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Aug 19 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"