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文献和实验ULTIMATE FREEZE-THAW LYSIS FOR MAMMALIAN CELLS
-abundance proteins (like Myc!).The protocol is described for harvesting mammalian cells from 6 or 10 cm tissue-culture dishes.A.CELL HARVEST.1.Remove media from cells.Wash in ice-cold PBS.2.Scrape cells in 1 ml of ice-cold PBS.Transfer to 1.5 ml microfuge
,and the cell pellets were frozen and stored at –80ºC. Lysis Lysis was accomplished with B-PER reagent supplemented with 1/50 vol 10-mg/ml chicken egg-white lysozyme (stored at –80ºC in aliquots),1/125 vol 5-mg/ml DNaseI (in 20 mM sodium acetate,pH adjusted
Chromatin Immunoprecipitation from Mammalian Cell Extracts
1.Crosslink protein DNA complexes in vivo Grow cells in suspension and collect 5 x108 cells by low speed centrifugation.Resuspend cells in 50ml media.In fume hood,add 1.4ml 37% fomaldehyde solution to a 50ml conical tube.Fill tube with cell culture
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