Anti-Caspase 3 Antibody, active (cleaved) form

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.

AB3623 detects only the cleaved p17 fragment and does not detect the precursor form in cells undergoing apoptosis.

Reactivity with other species has not been determined.

Cleavage specific synthetic peptide sequence (proprietary).

Epitope: active (cleaved) form

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).

Research Category
Apoptosis & Cancer

Metabolism

Research Sub Category
Caspases

Enzymes & Biochemistry

Western Blot Analysis:
1:100-1:200 (0.5-4 µg/mL) dilution of active apoptotic cells we used to detect Caspase-3.
Note when using tissue, levels of active caspase 3 can below detectable levels. Use CC119 (human caspase 3) incubated 30 minutes with 1 mM ATP (for activation) as a positive control on westerns. Alternatively, neuronal culture preparations incubated with staurosporine (0.5 µM, 10-18 hr) typically give a strong caspase 3 signal.

Immunohistochemistry:
1:10 dilution (10-20 µg/mL), AB3623 has reacted successfully in 4% PFA fixed, paraffin embedded tissues mouse tissues.

Immunohistochemistry(paraffin):
Optimal Staining of Caspase-3 Monoclonal Antibody: Mouse Small Intestine

17 kDa

Replaces: 04-440; 04-1088

ImmunoAffinity Purified

Purified rabbit polyclonal in buffer containing PBS, containing 30% glycerol, 0.5% BSA, 5 mM EDTA and 0.03% proclin.

Stable for 1 year at -20ºC from date of receipt.

Control
Tonsil and appendix tissue. Jurkat cells induced for apoptosis with recombinant soluble TRAIL and staurosporine treated mouse 3T3 cell lysates Skin/Tumor, Embryo.

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

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