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文献和实验Phosphoamino acid analysis:Mark Kamps's method
or enough to submerge your piece of membrane. 3.Incubate in 110℃ oven for 60 min. 4.Microfuge sample full speed for at least 1 min. 5.Transfer supernatant to new tube and lyophilize on Speedivac.(It takes about 3 hr.) 6.Resuspend in H2O,and microfuge 5 min.Spotting
Kamps's Western Blotting Protocol
from the shelf (Margie's).Stain for approximately 20 to 60 minutes.Rinse with water for 1 hr.Dry flat.Be careful.Buffers containing Tween 20 can reduce the binding of antiphosphotyrosine antibodies. Recipes Transfer buffer: Glycine 57.6 g Tris Base 12.0 g
C. elegans whole mount immunohistochemistry with Bouin's Fixative
pipette (600:200:10 Bouin's: MeOH: ßME also works). The tube is rocked on a Labquake shaker at RT for 30 minutes, then quick frozen in liquid N2. The tube is quickly thawed under running hot water until the solution melts (but before it warms up past RT
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