Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

SignalSilence® XIAP siRNA Kit (Human Specific) #6445

Kit Includes

SignalSilence® XIAP siRNA (Human Specific) #6446
pre-validated single sequence siRNA duplex guaranteed to specifically inhibit XIAP expression in specified cell lines.
SignalSilence® Control siRNA (Fluorescein Conjugate) #6201
a non-targeted negative control duplex designed to monitor transfection efficiency using microscopy and to monitor siRNA specificity.Kit contains enough for 4(S) or 10 (L) transfections.
Targeted XIAP Antibody #2042
to confirm silencing of XIAP expression.
Non-targeted Cofilin Antibody #3312
to control for protein loading and XIAP siRNA specificity.
*Transfection Reagent
designed specifically for highly efficient siRNA delivery in mammalian cells. Transfection reagent is supplied by Mirus.

*Store at 4° (packaged separately)

Western Blotting

Western Blotting

Western blot analysis of extracts from A549 cells 48 hours following mock transfection (-), transfection with non-targeted (control) siRNA or transfection with XIAP siRNA. XIAP was detected using XIAP Antibody #2042, and cofilin was detected using Cofilin Antibody #3312. The XIAP Antibody confirms silencing of XIAP expression, and the cofilin Antibody is used to control for protein loading and siRNA specificity.

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® XIAP siRNA Kit from Cell Signaling Technology allows the researcher to specifically inhibit XIAP expression. The kit utilizes 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 kits are rigorously tested in-house and have been shown to reduce protein expression in specified cell lines. SignalSilence® XIAP siRNA Kit includes XIAP siRNA, a target-specific XIAP Antibody to confirm the silencing of XIAP expression and a non-target antibody (Cofilin) to control for loading and monitor specificity of XIAP siRNA. Additionally, a fluorescein-labeled non-targeted siRNA control allows the user to monitor transfection efficiency and siRNA specificity. The kit also contains a transfection reagent designed solely for efficient siRNA delivery in mammalian cells.

Directions for Use

CST recommends transfection with 100 nM human specific XIAP siRNA. Decreased XIAP expression was seen 24-72 hours post-transfection. See Protocol for transfection procedure.

Background

The inhibitor of apoptosis protein (IAP) family consists of an evolutionarily conserved group of apoptosis inhibitors containing a conserved 70 amino acid BIR (baculovirus inhibitor repeat) domain (1,2). Human members of the family include c-IAP1, c-IAP2, XIAP, Surivivin, Livin and NAIP. Overexpression of IAP family members, particularly Survivin and Livin, in cancer cell lines and primary tumors suggest an important role for these proteins in cancer progression (3-5). In general, the IAP proteins function through direct interactions to inhibit the activity of several caspases, including caspase-3, caspase-7 and caspase-9 (5,6). In addition, binding of IAP family members to the mitochondrial protein Smac blocks its interaction with caspase-9, thereby allowing the processing and activation of the caspase (7).

Using RNA intereference, XIAP has been shown to function through MURR1 to regulate copper homeostasis (6).

  1. Deveraux, Q.L. and Reed, J.C. (1999) Genes Dev 13, 239-52.
  2. Deveraux, Q.L. et al. (1998) EMBO J 17, 2215-23.
  3. Altieri, D.C. et al. (1999) Lab Invest 79, 1327-33.
  4. Tamm, I. et al. (2000) Clin Cancer Res 6, 1796-803.
  5. Kasof, G.M. and Gomes, B.C. (2001) J Biol Chem 276, 3238-46.
  6. Deveraux, Q.L. et al. (1997) Nature 388, 300-4.
  7. Deveraux, Q.L. et al. (1998) EMBO J 17, 2215-23.
  8. Burstein, E. et al. (2004) EMBO J. 23, 244-254.

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.

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