Western blot analysis of cell extracts from Jurkat cells treated with 0.25 mg/ml cytochrome c, using Cleaved Caspase-3 (Asp175) Antibody #9661 without (left) and with (right) preincubation with cleavage-peptide.Learn more about how we get our images
Immunohistochemical staining of paraffin-embedded human tonsil, using Cleaved Caspase-3 (Asp175) Antibody #9661 preincubated with irrelevant control peptide (left) and with Cleaved Caspase-3 (Asp175) Blocking Peptide (right).Learn more about how we get our images
This peptide is used to block Cleaved Caspase-3 (Asp175) Antibody #9661 and #9664 reactivity.
The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Cleaved Caspase-3 (Asp175) Antibody #9661 by immunohistochemistry and Western Blotting.
Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry and Western blot protocols. For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 µl total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the relevant product data sheet.
For Western immunoblotting, add 10 µl of antibody and 10 µl of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room
temperature for 30 minutes before allowing to react with the blot.
Supplied in 20 mM potassium phosphate (pH 7.0), 50 mM NaCl, 0.1 mM EDTA, 1 mg/ml BSA, 5% glycerol and 1% DMSO. Store at –20°C.
Caspase-3 (CPP-32, Apoptain, Yama, SCA-1) is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins, such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP) (1). Activation of caspase-3 requires proteolytic processing of its inactive zymogen into activated p17 and p12 fragments. Cleavage of caspase-3 requires the aspartic acid residue at the P1 position (2).
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