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鼎国昌盛
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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
C. elegans whole mount immunohistochemistry with Bouin's Fixative
. If the worms remain yellow after an hr of washing, they usually end up impermeable to antibodies. Wash again with BTB and incubate an additional 2 or 3 hrs. Wash with BT (BTB lacking BME) one time, and AbA (1X PBS, 1.0% BSA, 0.5% Triton X-100, 10 mM NaAzide
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
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