Cell Signaling Technology

Product Pathways - MAPK Signaling

SignalSilence® HSP27 siRNA Kit (Human Specific) #6355

Kit Includes

SignalSilence® HSP27 siRNA (Human Specific) #6356
pre-validated single sequence siRNA duplex guaranteed to specifically inhibit HSP27 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 HSP27 (G31) Monoclonal Antibody #2402
to confirm silencing of HSP27 expression.
Non-targeted p42 MAP Kinase (3A7) Monoclonal Antibody #9107
to control for protein loading and HSP27 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 transfected with 100 nM control siRNA #6201 (-) or HSP27 siRNA (+), using HSP27 (G31) Monoclonal Antibody #2402 and p42 MAP Kinase (3A7) Monoclonal Antibody #9107. The HSP27 (G31) Monoclonal Antibody confirms silencing of HSP27 expression, and p42 MAP Kinase (3A7) Monoclonal Antibody is used to control for loading and specificity of HSP27 siRNA.

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® HSP27 siRNA Kit from Cell Signaling Technology allows the researcher to specifically inhibit HSP27 expression in human cell lines. 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. SignalSilence® HSP27 siRNA Kit includes HSP27 siRNA, a target-specific HSP27 Monoclonal Antibody to confirm the silencing of HSP27 expression and a non-target p42 MAP Kinase Monoclonal Antibody to control for loading and monitor specificity of HSP27 siRNA. Additionally, a fluorescent tagged non-targeted siRNA control allows the user to monitor transfection efficiency and siRNA specificity. The kit also includes a transfection reagent designed solely for efficient siRNA delivery in mammalian cells.

Directions for Use

CST recommends transfection with 100 nM human-specific HSP27 siRNA 48 to 72 hours prior to cell lysis. See Protocol for transfection procedure.

Background

Heat shock protein (HSP) 27 is one of the small HSPs that are constitutively expressed at different levels in various cell types and tissues. Like other small heat shock proteins, HSP27 is regulated at both the transcriptional and posttranslational levels (1). In response to stress, the expression level of HSP27 increases several-fold to confer cellular resistance to the adverse environmental change. HSP27 is phosphorylated at Ser15, Ser78 and Ser82 by MAPKAP kinase 2 as a result of the activation of the p38 MAP kinase pathway (2,3). Phosphorylation of HSP27 causes a change in its tertiary structure, which shifts from large homotypic multimers to dimers and monomers (4). It has been shown that phosphorylation and increased concentration of HSP27 modulates actin polymerization and reorganization (5,6).

Small interfering RNA (siRNA) has been used to specifically silence HSP27 expression in HeLa cells (7).

  1. Arrigo, A.P. and Landry, J. (1994) Cold Spring Harbor Laboratory Press, NY, 335-373.
  2. Landry, J. et al. (1992) J. Biol. Chem. 267, 794-803.
  3. Rouse, J. et al. (1994) Cell 78, 1027-1037.
  4. Rogalla, T. et al. (1999) J. Biol. Chem. 274, 18947-18956.
  5. Lavoie, J. et al. (1993) J. Biol. Chem. 268, 24210-24214.
  6. Rousseau, S. et al. (1997) Oncogene 15, 2169-2177.
  7. Park, K. et al. (2003) J. Biol. Chem. 278, 35272-35278.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

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.

Products