Rat pheochromocytoma-derived PC12 cells are an important cell model for studies of growth factor cell signaling pathways. PC12 has been used for a variety of studies where cell division and cell differentaiton have been induced through growth factor activation. Both NGF and FGF can promote neurite proliferation, while EGF and IGF-1 can induce mitosis in PC12 cells. In addition, PC12 cells can be used as a model of neuron development since they express many neuronal proteins and can differentiate into a neuron-like morphology.
PC12 cells grown in suspension cultures were serum starved for two hours and the cells were then either left untreated (Cat.# PL7091) or treated with NGF (200 ng/ml; PL7131) or Calyculin A (100 nM; Cat.# PL7101) or pervanadate (1 mM; Cat.# PL7111) for 30 minutes at 37°C. Cells were lysed in 1% SDS, 1.0 mM sodium ortho-vanadate, 1 mM sodium fluoride, 10 mM Tris (pH 7.4) buffer. Protein concentration was determined using the BCA method (Pierce) before diluting to final concentration and buffer.
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*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
This kit contains: