Product Pathways - Apoptosis / Autophagy
Cleaved Caspase-3 (Asp175) Blocking Peptide #1050
Description
This peptide is used to block Cleaved Caspase-3 (Asp175) Antibody #9661 and #9664 reactivity.
Quality Control
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.
Western Blotting
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.
IHC-P (paraffin)
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).
Applications
Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry and Western blot protocols.
Directions for Use
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.
Background
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 aspartic acid at the P1 position (2).
- Fernandes-Alnemri, T. et al. (1994) J. Biol. Chem. 269, 30761-30764.
- Nicholson, D. W. et al. (1995) Nature 376, 37-43.
Application References
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Companion Products
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
