Cell Signaling Technology

Product Pathways - Apoptosis

SignalSilence® Bcl-2 siRNA I #6441

Applications Reactivity
Transfection H (M) (R)

Reactivity Key:  H=Human  M=Mouse  R=Rat
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 (Fluorescein Conjugate) #6201 (-), SignalSilence® Bcl-2 siRNA I (+) or SignalSilence® Bcl-2 siRNA II #6516 (+), using Bcl-2 (50E3) Rabbit mAb and α-Tubulin (11H10) Rabbit mAb #2125. Bcl-2 (50E3) rabbit mAb confirms silencing of Bcl-2 expression, while the α-tubulin (11H10) rabbit mAb is used to control for loading and specificity of Bcl-2 siRNA.

Description

SignalSilence® Bcl-2 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit Bcl-2 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.

Directions for Use

CST recommends transfection with 100 nM SignalSilence® Bcl-2 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

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 anti-apoptotic functions (7).

Silencing Bcl-2 expression by RNA interference induces p53 dependent apoptosis (8).

  1. Murphy, K.M. et al. (2000) Cell Death Differ 7, 102-11.
  2. Zhu, L. et al. (1999) J Biol Chem 274, 33267-73.
  3. Maundrell, K. et al. (1997) J Biol Chem 272, 25238-42.
  4. Yamamoto, K. et al. (1999) Mol Cell Biol 19, 8469-78.
  5. Ling, Y.H. et al. (1998) J Biol Chem 273, 18984-91.
  6. Huang, S.T. and Cidlowski, J.A. (2002) FASEB J 16, 825-32.
  7. Deng, X. et al. (2001) J Biol Chem 276, 23681-8.
  8. Jiang, M. and Milner, J. (2003) Genes Dev 17, 832-7.

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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