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Technical Data Sheet

PL7101

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PC12 + Calyculin A
Lysate
Price
Size

$75
100 μl

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Enlarge

Western blot of rat PC12 cells untreated (lanes 1 & 3) or treated with calyculin A (lanes 2 & 4). The blot was probed with anti-CRMP2 (C-terminal Region) antibody (lanes 1 & 2) or anti-CRMP2 (Thr-555) antibody (lanes 3 & 4).

Application
Dilution


WB
20 μl/lane



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
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 Calyculin A (100 nM; Cat.# PL7101) 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.

Buffer and Storage
Cell Lysates are supplied at a concentration of 1 mg/ml in electrophoresis sample buffer (62.5 mM Tris pH 6.8, 2% SDS, 5% glycerol, 0.003% bromophenol blue, 0.9% β-mercaptoethanol). Store at –20°C. Do not boil or dilute. Stable for 1 year.
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PC12 (NGF-differentiated) Lysate

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