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文献和实验- AT supplement : 50X stock , Sigma A-7422. (Store at -20°C.) - Kanamycin Sulfate: 100X from Gibco-BRL 600-5160AG . (Store aliquots at -20° C) - MCM: Macrophage Conditioned Medium. (used
HIS-tagged protein purification
1) grow 20ml cells O/N 37 C, dilute 50X into prewarmed LB, grow to 0.6 OD or about 1hr. Induce w/ 2mM IPTG (238mg/0.5l), grow 3hr, spin 6k GS3 10', freeze at -70 ℃.2) resuspend cells (from 500ml) in fliptop in 25 mls. UPB8.0, freeze lN2.3) thaw
GST fusion protein purification GST融合蛋白纯化
1)grow 20ml cells O/N37℃ dilute 50X into prewarmed LB,grow to 0.6 OD or about 1hr.Induce w/ 2mM IPTG (238mg/0.5l),grow 3hr,spin 6k GS3 10',freeze at 70℃2)extract cells (from 500ml)in 25 mls.Heintz Buffer plus triton (HBT)by gentle pipette
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