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 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.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
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.
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Read what people are saying who have used Human Caspase-3 Antibody.
We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Western Blot.
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.