Cell Signaling Technology

Product Pathways - Apoptosis

SignalSilence® Caspase-10 siRNA I #6357

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, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® Caspase-10 siRNA I (+), using Caspase-10 Antibody #9752 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The Caspase-10 Antibody confirms silencing of Caspase-10 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.

Description

SignalSilence® Caspase-10 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Caspase-10 expression using RNA interference, a method whereby 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 target protein expression by western analysis.

Quality Control

Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.

Directions for Use

CST recommends transfection with 100 nM Caspase-10 siRNA I 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.

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-10 is a DED (death effector domain)-containing caspase and functions as an initiator caspase in Fas/TNF induced apoptosis (1). Four isoforms of caspase-10 have been identified: caspase-10a (Mch4), caspase-10b (FLICE2), caspase-10c and caspase-10d. They have the same prodomain but different mature large and small subdomains (2-4). Upon death ligand-receptor binding, caspase-10 is coupled to the multimeric Fas/TNF receptor complex via DED/FADD adaptor interaction (1-4). This complex processes procaspase-10 into a large active fragment and a small fragment. Cleaved caspase-10 further processes other caspase members, including caspase-3 and caspase-7, to initiate a caspase cascade, leading to apoptosis (3-6).

  1. Nunez, G. et al. (1998) Oncogene 17, 3237-3245.
  2. Ng, P. W. et al. (1999) J. Biol. Chem. 274, 10301-10308.
  3. Vincenz, C. and Dixit, V.M. (1997) J. Biol. Chem. 272, 6578-6583.
  4. Fernandez-Alnemri, T. et al. (1996) Proc. Natl. Acad. Sci. USA 93, 7464-7469.
  5. Srinivasula, S. M. et al. (1996) Proc. Natl. Acad. Sci. USA 93, 14486-14491.
  6. Wang, J. et al. (1999) Cell 98, 47-58.

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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