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Chen
et al. Vet Res (2021) 52:73
bounding to protein for 60 min at 4 ℃. Then the beads
were washed five times with buffer A [150 mM KCl,
25 mM Tris pH 7.4, 5 mM EDTA, 0.5 mM DTT, 0.5%
NP40, 1 mM PMSF, 100 U/mL SUPERAsin and 1 × pro-
tease inhibitor cocktail (Sigma)]. After rotating at 1000
g
for 5 min, the precipitate
was added with the protein
loading buffer and loaded to precast 12% gradient Bis–
Tris gel for further analysis
by Coomassie brilliant blue
staining. The different strips were
then cut for mass spec-
trometry analysis.
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