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Analysis Note

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

Application

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

Research CategoryApoptosis & CancerMetabolism

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 Sub CategoryCaspasesEnzymes & Biochemistry

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

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.

Immunogen

Epitope: active (cleaved) form

Cleavage specific synthetic peptide sequence (proprietary).

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: 04-440; 04-1088

Other Notes

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

Physical form

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

ImmunoAffinity Purified

Specificity

Reactivity with other species has not been determined.

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

Storage and Stability

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

Target description

17 kDa