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文献和实验DETECTION OF ß-GALACTOSIDASE AND ALKALINE PHOSPHATASE ACTIVITIES IN TISSUE
EGTA stock, pH 8.0) in 100 ml PBS, pH 7.2-7.4 Heat about 80ml H2 O to 600 C and add paraformaldehyde; add 1-2 drops of 10M NaOH to get paraformaldehyde in solution. Cool to room temperature, add 10 ml 10X PBS, adjust pH with HCl, add MgCl
Metabolic labeling Immunoprecipitation
•Expre 35 S35 S Protein labeling mix (NEN,#NEG-072),7.9mCi/ml • Transfection reagents: CaCl2 ,x2HBS,DNA •Labeling Medium: RPMI1640(Met,Cys-free; Sigma R-7513)supplemented w/ 10% dialyzed FCS,& Gln • Chase Medium: Labeling medium (above)+ 500µg
High efficiency LiAc transformation
media and incubate with shaking overnight at 30o C. 2. Count o/n culture and inoculate 50 mls of warm YPED to a cell density of 5 x 106 /ml culture. Note: i) Dilute overnight cultures 10-1 or more in water. ii) Carefully place 10
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