Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

SignalSilence® Bcl-2 siRNA Kit (Human Specific) #6440

Kit Includes

SignalSilence® Bcl-2 siRNA (Human Specific) #6441
pre-validated single sequence siRNA duplex guaranteed to specifically inhibit Bcl-2 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.
Bcl-2 Antibody #2872
to confirm silencing of Bcl-2.
Non-targeted p42 MAPK Antibody #9108
to control for protein loading and Bcl-2 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 HeLa cells 48 hours following mock transfection, transfection with nonspecific (control) siRNA or transfection with Bcl-2 siRNA. Bcl-2 was detected using Bcl-2 Antibody #2872 and p42 was detected using p42 MAPK Antibody #9108. The Bcl-2 Antibody confirms silencing of Bcl-2 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® Bcl-2 siRNA Kit (Human Specific) from Cell Signaling Technology allows the researcher to specifically inhibit Bcl-2 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® Bcl-2 siRNA Kit (Human Specific) includes Bcl-2 siRNA, a target-specific Bcl-2 Antibody to confirm the silencing of Bcl-2 expression and a non-target antibody (p42 MAPK) to control for loading and monitor specificity of Bcl-2 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 Bcl-2 siRNA. Decreased Bcl-2 expression was observed 24-72 hours post-transfection. See Protocol for transfection procedure.

Background

Bcl-2 exerts a survival function in response to a wide range of apoptotic stimuli through inhibition of mitochondrial cytochrome c release (1). It has been implicated in modulating mitochondrial calcium homeostasis and proton flux (2). Several phosphorylation sites have been identified within Bcl-2 including Thr56, Ser70, Thr74 and Ser87 (3). It has been suggested that these phosphorylation sites may be targets of the ASK1/MKK7/JNK1 pathway, and that phosphorylation of Bcl-2 may be a marker for mitotic events (4,5). Mutation of Bcl-2 at Thr56 or Ser87 inhibits its anti-apoptotic activity during glucocorticoid-induced apoptosis of T lymphocytes (6). Interleukin 3 and JNK-induced Bcl-2 phosphorylation at Ser70 may be required for its enhanced antiapoptotic functions (7).

Silencing Bcl-2 expression by RNA interfence induces p53-depedent apoptosis (8).

  1. Murphy, K.M. et al. (2000) Cell Death Differ. 7, 102-111.
  2. Zhu, L. et al. (1999) J. Biol. Chem. 274, 33267-33273.
  3. Maundrell, K. et al. (1997) J. Biol. Chem. 272, 25238-25242.
  4. Yamamoto, K. et al. (1999) Mol. Cell. Biol. 19, 8469-8478.
  5. Ling, Y.H. et al. (1998) J. Biol. Chem. 273, 18984-18991.
  6. Huang, S.J. and Cidlowski, J.A. (2002) FASEB J. 16, 825-832.
  7. Deng, X. et al. (2001) J. Biol. Chem. 276, 23681-23688.
  8. Jiang, M. and Milner, J. (2003) Genes Dev. 17(7) , 832-837.

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.

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