小中大材料：古菌
提取方法： cells were harvested from a 100 mL culture and
washed twice in 3 volumes w/v 1.5 M Tris-HCl buffer at
pH 7.1. Cells were resuspended in 3 volumes w/v 50 mM
Tris-HCl buffer at pH 7.1 supplemented with 0.8% SDS and
phosphatase and protease inhibitor cocktails according to the
supplier.cells homogenized by sonication,the lysate
was centrifuged (15 600 3 g, 20 min, 47C). The supernatant
was boiled (5 min), chilled on ice (5 min), and centrifuged
(15 600 3 g, 30 min, 47C). The sample was resuspended in 0.5 mL 250 mM glycerol
supplemented with 10 mM TEA prior to dilution.
上样量：300ug
IEF：Step1 30 S time: 10:00
Step2 200 G time: 02:00
Step3 500 G time: 02:00
Step4 1000 G time: 01:00
Step5 5000 G time: 01:00
Step6 10000 G time: 01:00
Step7 10000 S Vh: 60000
Step8 1000 S time: 01:00
