Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-pro Caspase-3 antibody [E83-103] - BSA and Azide free (ab238440)

Overview

  • Product name

    Anti-pro Caspase-3 antibody [E83-103] - BSA and Azide free
    See all pro Caspase-3 primary antibodies
  • Description

    Rabbit monoclonal [E83-103] to pro Caspase-3 - BSA and Azide free
  • Host species

    Rabbit
  • Specificity

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

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

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human pro Caspase-3 (N terminal). The exact sequence is proprietary. A synthetic peptide corresponding to residues following Ser29 of human caspase-3 (N-terminus of p17 subunit).

  • Positive control

    • WB: Wild-type HAP1 whole cell lysate. IHC-P: Human colon adenocarcinoma tissue. ICC/IF: HeLa cells. Flow Cyt: Jurkat cells.
  • General notes

    ab238440 is the carrier-free version of ab32499 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab238440 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this 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.

    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

    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E83-103
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab238440 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
IHC-P Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 35 kDa (predicted molecular weight: 31 kDa).
Flow Cyt Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

Target

  • Relevance

    Caspases 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 localization

    Cytoplasmic
  • 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

Images

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab32499).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab32499)

  • ICC/IF image of ab32499 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

    The cells were fixed in 4% formaldehyde (10 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32499, 5 µg/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 1.43 µM.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab32499)

  • Overlay histogram showing Jurkat (Human T cell leukemia cell line from peripheral blood) 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) (1 µg/1 x 106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab32499)

References

ab238440 has not yet been referenced specifically in any publications.

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