Cell Signaling Technology

Product Pathways - Apoptosis

SignalSilence® Caspase-3 siRNA I #6466

Applications Reactivity
Transfection H

Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells 48 hours following mock transfection, transfection with non-targeted (control) siRNA or transfection with Caspase-3 siRNA (100 nM). Caspase-3 was detected using Caspase-3 Rabbit Monoclonal Antibody #9665, and p42 MAPK was detected using p42 MAPK Antibody #9108.

Fluorescent Detection

Fluorescent Detection

Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency.

Description

SignalSilence® Caspase-3 siRNA allows the researcher to specifically inhibit Caspase-3 expression using RNA interference, a method in which gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce protein expression in specified cell lines.

Directions for Use

CST recommends transfection with 100 nM Caspase-3 siRNA. Decreased Caspase-3 expression was observed 24-72 hours post-transfection. See Protocol for transfection procedure.

Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.

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 the aspartic acid residue at the P1 position (2).

In addition to its central role in apoptosis, Caspase-3 has been shown to play a role in erythroid differentiation through the use of RNA interference (3).

  1. Fernandes-Alnemri, T. et al. (1994) J. Biol. Chem. 269, 30761-30764.
  2. Nicholson, D. W. et al. (1995) Nature 376, 37-43.
  3. Carlile, G. W. et al. (2004) Blood 103, 4310-4316.

Application References

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Companion Products

Limited Use Label License, RNA interference: This product is licensed under European Patent 1144623 and foreign equivalents from Ribopharma AG, Kulmbach, Germany and is provided only for use in non-commercial research specifically excluding use (a) in drug discovery or drug development, including target identification or target validation, by or on behalf of a commercial entity, (b) for contract research or commercial screening services, (c) for the production or manufacture of siRNA-related products for sale, or (d) for the generation of commercial databases for sale to Third Parties. Information about licenses for these and other commercial uses is available from Ribopharma AG, Fritz-Hornschuch-Str. 9, D-95326 Kulmbach, Germany.

For Research Use Only. Not For Use In Diagnostic Procedures.

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