Caspase-3 (CPP32, Apopain) is the most extensively studied apoptotic protein. Caspase-3 is synthesized as an inactive proenzyme (32 kDa) that is processed in cells undergoing apoptosis by self-proteolysis and/or cleavage by another upstream protease. The processed form of caspase-3 consists of large (17 kDa) and small (12 kDa) subunits which associate to form an active enzyme. The active caspase-3 proteolytically cleaves and activates other caspases, as well as relevant targets in the cells such as PARP and DFF.

Anti-Caspase 3, active (cleaved) is a rabbitpolyclonal antibody that detects active Caspase 3. It targets an epitope within13 amino acids from the C-terminal region.

KLH-conjugated linear peptide corresponding to 13 amino acids from theC-terminal region of human active Caspase 3.

Anti-Caspase 3 Antibody, active (cleaved) form detects level of Caspase 3 and has been published and validated for use in Immunofluorescence (IF), Immunohistochemistry (IHC), and Western Blot (WB).

Quality Control Testing

Evaluated by Western Blotting in Jurkat cells treatedwith Camptothecin. Western Blotting Analysis (WB): A 1:500 dilution from a representative lot detectedactive Caspase-3 in lysate from Jurkat cells treated with Camptothecin (1 μM; 3 hours).

Tested Applications Immunohistochemistry (Paraffin) Analysis (IHC): A 1:200dilution from a representative lot detected Caspase 3, active (cleaved) inhuman tonsil and human colon tissuesections. Immunocytochemistry Analysis (ICC): A 1:500 dilution froma representative lot detected active Caspase-3 in HeLa cells treated withStaurosporine (0.5 μM for 6 hours.). Immunocytochemistry Analysis (ICC): A 1:500 dilution froma representative lot detected active Caspase-3 in HeLa cells treated withCamptothecin (3 μM for 6 hours.). Peptide Inhibition Assay (PIA): Target band detection inlysate from Jurkat cells treated with Camptothecin (1μM; 3 hours)was prevented by pre-blocking of a representative lot with the immunogenpeptide. Note: Actual optimal working dilutions must bedetermined by end user as specimens, and experimental conditions may vary withthe end user.

Target molecular weight ~17 kDa observed. 17kDa calculated. Uncharacterized bands may be observed in some lysate(s).

Replaces: 04-440; 04-1088

Purified rabbit polyclonal antibody in PBS with 0.05% sodium azide with 30% glycerol.

Store at -10°C to -25°C. Handling Recommendations: Upon receipt andprior to removing the cap, centrifuge the vial and gently mix the solution.Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thawcycles, which may damage IgG and affect product performance.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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