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上海希言科学仪器有限公司
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文献和实验7.5 5ml of 1M 0.3M NaCl150ml 1M or 60ml 2.5M 1mM EDTA 1ml of 0.5M 0.02% NaN3 1ml of 10% CREB Column- Purification Trial #1 Sample #5-C4-1-1 (pg.25)lysed in 4.5ml (3 vol.)0.75M KCl+ Buffer C (188x106cells) Thawed on ice. Pellet 20min.in microfuge
Bases Buffer is Buffer C (as per Schroter)(as per SRF)pg.27 20mM Hepes pH 7.9 0.1% NP40 0.2mM EDTA 20% glycerol Lysis Buffer: 0.75M KCl Need buffers containing 0,0.1M,0.6M & 1.0M KCL in addition to 0.3M,1.5M & 2.0 used in SRF purification pg
lysis buffer (high salt) ) 2 x 1 ml CHIP wash buffer 2 x 1 ml TE elute immunoprecipitations: add 75 µl elution buffer incubate for 10 min at 65 °C spin, take supernatant, elute pellet again with 75 µl
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