Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

SignalSilence® Bax siRNA Kit #6320

Kit Includes

SignalSilence® Bax siRNA (Human Specific) #6321
pre-validated single sequence siRNA duplex guaranteed to specifically inhibit Bax 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 Bax antibody #2772
to confirm silencing of Bax expression.
Non-targeted p42 MAP Kinase Antibody #9108
to control for protein loading and Bax 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 24 hours following mock transfection, transfection with non-targeted (control) siRNA or transfected with Bax siRNA. Bax was detected using Bax Antibody #2772, and p42 was detected using p42 MAPK Antibody #9108. The Bax Antibody confirms silencing of Bax 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® Bax siRNA Kit from Cell Signaling Technology allows the researcher to specifically inhibit Bax 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. SignalSilence® Bax siRNA Kit includes Bax siRNA, a target-specific Bax Antibody to confirm the silencing of Bax expression and a non-target antibody (p42 MAPK) to control for loading and monitor specificity of Bax 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 Bax siRNA. Decreased Bax expression seen 24 hours post-transfection. See Protocol for transfection procedure.

Background

Bax is a key component for cellular induced apoptosis through mitochondrial stress (1). Upon apoptotic stimulation, Bax forms oligomers and translocates from the cytosol to the mitochondrial membrane (2). Through interactions with pore proteins on the mitochondrial membrane, Bax increases the membrane's permeability, which leads to the release of cytochrome c from mitochondria, activation of caspase-9 and initiation of the caspase activation pathway for apoptosis (3,4).

Bax expression was effectively reduced in T lymphocytes by RNA interference, demonstrating the dependence on Bax to trigger the release of AIF from the mitochondria during apoptosis (5).

  1. Wei, M.C. et al. (2001) Science 292, 727-730.
  2. Jurgensmeier, J.M. et al. (1998) Proc. Natl. Acad. Sci. USA 95, 4997-5002.
  3. Narita, M. et al. (1998) Proc. Natl. Acad. Sci. USA 95, 14681-14686.
  4. Marzo, I. et al. (1998) Science 281, 2027-2031.
  5. Bidere, N. et al. (2003) J. Biol. Chem. 278(33) , 31401-31411.

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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