Cell Signaling Technology

Product Pathways - Apoptosis / Autophagy

SignalSilence® c-Myc siRNA Kit #6340

Kit Includes

SignalSilence® c-Myc siRNA (Human Specific) #6341
pre-validated single sequence siRNA duplex guaranteed to specifically inhibit c-Myc 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 c-Myc Antibody #9402
to confirm silencing of c-Myc expression.
Non-targeted p42 MAPK Antibody #9108
to control for protein loading and c-Myc 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 c-Myc siRNA. c-Myc was detected using c-Myc Antibody #9402, and p42 was detected using p42 MAPK Antibody #9108. The c-Myc Antibody confirms silencing of c-Myc 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® c-Myc siRNA Kit from Cell Signaling Technology allows the researcher to specifically inhibit c-Myc 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® c-Myc siRNA Kit includes c-Myc siRNA, a target-specific c-Myc Antibody to confirm the silencing of c-Myc expression and a non-target antibody (p42 MAPK) to control for protein loading and monitor specificity of c-Myc 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 c-Myc siRNA. Decreased c-Myc expression was observed 24-72 hours post-transfection. See Protocol for transfection procedure.

Background

Members of the Myc/Max/Mad network function as transcriptional regulators with roles in various aspects of cell behavior including proliferation, differentiation and apoptosis (1). These proteins share a common basic-helix-loop-helix leucine zipper (bHLH-ZIP) motif required for dimerization and DNA-binding. Max was originally discovered based on its ability to associate with c-Myc and found to be required for the ability of Myc to bind DNA and activate transcription (2). Subsequently, Max has been viewed as a central component of the transcriptional network, forming homodimers as well as heterodimers with other members of the Myc and Mad families (1). The association between Max and either Myc or Mad can have opposing effects on transcriptional regulation and cell behavior (1). The Mad family consists of four related proteins; Mad1, Mad2 (Mxi1), Mad3 and Mad4, and the more distantly related members of the bHLH-ZIP family, Mnt and Mga. Like Myc, the Mad proteins are tightly regulated with short half-lives. In general, Mad family members interfere with Myc-mediated processes such as proliferation, transformation and prevention of apoptosis by inhibiting transcription (3,4).

siRNA has been used previously to inhibit the expression of c-Myc in the human colon tumor cell line, HTC116 (6).

  1. Baudino, T.A. and Cleveland, J.L. (2001) Mol. Cell. Biol. 21, 691-702.
  2. Blackwood, E.M. and Eisenman, R.N. (1991) Science 251, 1211-1217.
  3. Henriksson, M. and Luscher, B. (1996) Adv. Cancer Res. 68, 109-182.
  4. Grandori, C. et al. (2000) Annu. Rev. Cell Dev. Biol. 16, 653-699.
  5. Williams, N. S. et al. (2003) Clin. Cancer Res. 9, 931-946.

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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