Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency.
Western blot analysis of extracts from HeLa cells, transfected with control (-) or cdc2 (+) siRNA. cdc2 was detected using cdc2 (POH1) Monoclonal Antibody#9116, and p42 was detected using p42 MAPK (3A7) Monoclonal Antibody #9107. cdc2 (POH1) Monoclonal Antibody confirms silencing of cdc2 expression, and p42 MAPK (3A7) Monoclonal Antibody is used to control for loading and specificity of cdc2 siRNA.
CST recommends transfection with 50-100 nM cdc2 siRNA 48 hours prior to cell lysis. See protocol for transfection procedure.
SignalSilence® siRNA is supplied in RNAse-free water. Aliquot and store at -20ºC.
SignalSilence® cdc2 siRNA from Cell Signaling Technology allows the researcher to specifically inhibit cdc2 expression using 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 products are rigorously tested in-house and have been shown to reduce protein expression in specified cell lines.
The entry of eukaryotic cells into mitosis is regulated by cdc2 kinase activation, a process controlled at several steps including cyclin binding and phosphorylation of cdc2 at Thr161 (1). However, the critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of cdc2 at Thr14 and Tyr15 (2). Phosphorylation at Thr14 and Tyr15, resulting in inhibition of cdc2, can be carried out by Wee1 and Myt1 protein kinases (3,4). The cdc25 phosphatase may be responsible for removal of phosphates at Thr14 and Tyr15 and subsequent activation of cdc2 (1,5).
RNA interference has been used to specifically silence cdc2 expression in HeLa cells, resulting in cell cycle arrest prior to spindle formation (7).
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