Product Pathways - Cell Cycle / Checkpoint
SignalSilence® Chk1 siRNA (Human Specific) #6241
Western Blotting
Western blot analysis of extracts from HeLa cells transfected with non-targeted (-) or targeted (+) siRNA. Chk1 was detected using the Chk1 Antibody #2345, and p42 was detected using the p42 MAPK Antibody #9108. The Chk1 Antibody confirms silencing of Chk1 expression, and the p42 MAPK Antibody was used to control for loading and specificity of Chk1 siRNA.
Fluorescent Detection
Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency.
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).
- Martinho, R.G. et al. (1998) EMBO J. 17, 7239-7249.
- Zhao, H. et al. (2001) Mol. Cell. Biol. 21, 4129-4139.
- Zeng, Y. et al. (1998) Nature 395, 507-510.
- Shieh, S. et al. (2000) Genes Dev. 14, 289-300.
- 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!
Companion Products
- 6240 SignalSilence® Chk1 siRNA Kit (Human Specific)
- 2345 Chk1 Antibody
- 6201 SignalSilence® Control siRNA (Fluorescein Conjugate)
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
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.
