Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Knockout Validated, Western Blot

Cited:

Immunohistochemistry-Paraffin, Western Blot, Immunoprecipitation, ELISA-Based Protein Pin Array

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 84803
View all Caspase-3 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Caspase-3 (18 kDa subunit)
aa 29-175
Accession # P42574

Specificity

Detects human precursor Caspase-3 and the 18 kDa subunit generated during apoptosis.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human Caspase‑3 Antibody

Detection of Human Precursor Caspase-3 and p18 Subunit antibody by Western Blot.

Detection of Human Precursor Caspase‑3 and p18 Subunit by Western Blot.

Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 µM staurosporine (STS) for 3 hours. PVDF membrane was probed with 1 µg/mL of Human Caspase-3 Monoclonal Antibody (Catalog # MAB707), followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Specific bands were detected for precursor Caspase-3 and the p18 subunit at approximately 36 and 18 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
Western Blot Shows Human Caspase-3 Antibody Specificity by Using Knockout Cell Line.

Western Blot Shows Human Caspase‑3 Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Caspase-3 knockout HeLa cell line (KO). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Caspase-3 Monoclonal Antibody (Catalog # MAB707) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Caspase-3 at approximately 32 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Applications for Human Caspase‑3 Antibody

Application
Recommended Usage

Knockout Validated

Caspase‑3 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in Caspase‑3 knockout HeLa cell line.

Western Blot

1 µg/mL
Sample: Staurosporine-treated Jurkat human acute T cell leukemia cell line

Reviewed Applications

Read 1 review rated 5 using MAB707 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from ascites

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Caspase-3

Caspase-3 (Cysteine-aspartic acid protease 3/Casp3; also Yama, apopain and CPP32) is a 29 kDa heterodimer that belongs to the peptidase C14A family of enzymes. It is widely expressed, and considered to be the major executioner caspase in the apoptotic cascade. Human procaspase-3 is a 32 kDa, 277 amino acid (aa) protein and is normally an inactive homodimer. Following cell stress/activation, procaspase-3 undergoes proteolysis to generate an N-terminal 148 aa p17/17 kDa subunit (aa 29-175), plus a 102 aa C-terminal p12/12 kDa subunit. These subunits noncovalently heterodimerize, and associate with another p17/p12 heterodimer to form an active enzyme. There is one potential variant that shows an alternative start site nine aa upstream of the standard start site coupled with a 21 aa substitution for aa 162-277. Over aa 29-175, human and mouse caspase-3 share 87% aa identity.

 

Alternate Names

Apopain, CASP3, Caspase3, CPP32, LICE-1, YAMA

Entrez Gene IDs

836 (Human); 12367 (Mouse)

Gene Symbol

CASP3

UniProt

P42574 (Rabbit)

Additional Caspase-3 Products

Product Documents for Human Caspase‑3 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Caspase‑3 Antibody

For research use only

Related Research Areas

Cysteine Proteases and Regulators

Citations for Human Caspase‑3 Antibody

Customer Reviews for Human Caspase‑3 Antibody (1)

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  • Human Caspase-3 Antibody
    Name: David Timm
    Application: Western Blot
    Sample Tested: Blood erythrocytes
    Species: Human
    Verified Customer | Posted 02/28/2017
    The antibody works fine for what it is, but I think it will not work for our experiments. Compared to Jurkat or other cell types you only get very faint bands for caspase in RBCs unless you fix your membrane after the transfer, this gave a false positive comparing a negative and positive control for caspase activation. We had to switch to measuring samples only with flow. If your experiments are compatible it's a good antibody though. We tried this experiment with several antibodies of different companies and this one did give a nicer looking band.
    Human Caspase‑3 Antibody MAB707

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