Product Pathways - Cell Cycle / Checkpoint

SignalSilence® Chk1 siRNA Kit (Human Specific) #6240

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Kit Includes

SignalSilence^® Chk1 siRNA (Human Specific) #6241

pre-validated single sequence siRNA duplex guaranteed to specifically inhibit Chk1 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.

Targeted Chk1 Antibody #2345

to confirm silencing of Chk1 expression.

Non-targeted p42 MAPK Antibody #9108

to control for protein loading and Chk1 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)

Description

SignalSilence® Chk1 siRNA Kit from Cell Signaling Technology allows the researcher to specifically inhibit Chk1expression. The kit utilizes RNA interference, a method in which gene expression can be selectivelysilenced 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® Chk1 siRNA Kit includes a target-specific Chk1 Antibody to confirm the silencing of Chk1 expression and a non-target antibody (p42 MAPK) to control for loading and monitor specificity of Chk1 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 50 nM Chk1 siRNA 24 hours prior to cell lysis. See protocol for transfection procedure.

Background

Chk1 kinase acts downstream of ATM/ATR kinase to play an important role in DNA damage checkpoint control, embryonic development and tumor suppression (1). Activation of Chk1 involves phosphorylation of Ser317 and Ser345 and occurs in response to blocked DNA replication and certain forms of genotoxic stress (2). Chk1 is also phosphorylated at Ser280 and Ser296 following DNA damage. Activated Chk1 can inactivate cdc25C via phosphorylation at Ser216, blocking the activation of cdc2 and transition into mitosis (3). Chk1 can also phosphorylate p53 at Ser20 in vitro (4).

Small Interfering RNA (siRNA) has been used to specifically silence chk1 in various human cancer cell lines (5).

1. Martinho, R.G. et al. (1998) EMBO J. 17, 7239-7249.
2. Zhao, H. et al. (2001) Mol. Cell. Biol. 21, 4129-4139.
3. Zeng, Y. et al. (1998) Nature 395, 507-510.
4. Shieh, S. et al. (2000) Genes Dev. 14, 289-300.
5. Chen, Z. et al. (2003) Mol. Cancer Ther. 2(6) , 543-548.

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!

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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.