赛默飞世尔Thermo Fisher X100 Membra

原创：western protocol

of a 10% gel 4 ml H2O 3.3ml 30% acrylamide/bisacrylamide (29:1 mix) 2.5 ml1.5M Tris(PH 8.8) 10% SDS 100 µl 10% ammonium persulfate 100 µl TEMED 8 µl 2.  Stacking gel: 4 ml 2.7ml water 1 ml 0.67ml 30% acrylamide/bisacrylamide (29:1 mix) 0.ml 1.0M

v468 chapter 5 糖原合成酶激酶-3 的抑制

for cell lysis: 10 mM Tris-HCl, pH 7.4,50 mM NaCl, 5 mM EDTA, 50 mM NaF, 30 mM Na 4 P 2 O 7 ,and 1% (v/v) Triton-X100. 5. Protease inhibitors and phosphatase inhibitor are added fresheach time with the following final concentrations: 10 μg/mLaprotinin

WESTERN PROTOCOL

For 500,000 cells, lyse with 20 µl. Spin at 14,000 rpm (16,000 g) in an Eppendorf microfuge for 10 min at 4°C. Transfer the supernatant to a new tube and discard the pellet. Determine the protein concentration (Bradford assay, A280