Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

SignalSilence® Survivin siRNA Kit #6350

Kit Includes

SignalSilence® Survivin siRNA #6351
pre-validated single sequence siRNA duplex guaranteed to specifically inhibit survivin 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.
Survivin (6E4) Monoclonal Antibody #2802
to confirm silencing of survivin expression.
Non-targeted p42 MAPK Antibody #9108
to control for protein loading and survivin siRNA specificity.
*Transfection Reagent
designed specifically for highly efficient siRNA delivery in mammalian cells. Transfection reagent is supplied by Mirus.

*Store at 4°C (packaged separately)

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells 48 hours following mock transfection, transfection with nonspecific (control) siRNA or transfection with Survivin siRNA. Survivin was detected using Survivin (6E4) Monoclonal Antibody #2802, and p42 was detected using p42 MAPK Antibody #9108. The Survivin Antibody confirms silencing of Survivin expression, and the p42 MAPK 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® Survivin siRNA Kit from Cell Signaling Technology allows the researcher to specifically inhibit survivin 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® Survivin siRNA Kit includes Survivin siRNA, a target-specific Survivin Antibody to confirm the silencing of survivin expression and a non-target antibody (p42 MAPK) to control for loading and monitor specificity of Survivin 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 Survivin siRNA. Decreased survivin expression was observed 24-72 hours post-transfection. See Protocol for transfection procedure.

Background

Survivin is a 16 kDa anti-apoptotic protein highly expressed during fetal development and cancer cell malignancy (1). Survivin binds and inhibits caspase-3, controlling the checkpoint in the G2/M-phase of the cell cycle through inhibiting apoptosis and promoting cell division (2,3). This regulatory process requires the phosphorylation of survivin at Thr34 by p34 cdc2 kinase (4). Gene targeting using a Thr34 phosphorylation-defective survivin mutant, as well as antisense survivin, has been shown to inhibit tumor growth (5,6).

Survivin expression has been specifically silenced using siRNA in HeLa cells (7).

  1. Reed, J.C. and Reed, S.I. (1999) Nature Cell Biol. 1, 199-200.
  2. Li, F. et al. (1998) Nature 396, 580-584.
  3. Li, F. et al. (1999) Nat. Cell Biol. 1, 461-466.
  4. O'Connor, D.S. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 13103-13107.
  5. Olie, R.A. et al. (2000) Cancer Res. 60, 2805-2809.
  6. Grossman, D. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 635-640.
  7. Carvalho, A. et al. (2003) J. Cell Sci. 116, 2987-2998.

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.

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.

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