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文献和实验Metabolic labeling Immunoprecipitation
survey meter.Wipe test + LSC is necessary. Chemical properties... Use either 35S-methionine or cysteine or both.Recommend using Expre 35 S35 S Protein labeling mix (NEN).Before opening the cap of the vial,thaw the solution completely and open the cap
Metabolic labeling Immunoprecipitation
cells twice with Labeling Medium. 2. Add 1.5ml/well Labeling Medium 3. Add 20-30µl (~250µCi) of [35 S] methionine/cysteine /well 4. Culture 1h at 37°C (pulse) 5. Add 1.5ml/well Chase Medium 6. Culture o/n (chase) 7. Harvest metabolically
Northern Blotting方法(Howell Lab)
inverted because the RNA pellets on the bottom. (DNA can be recovered from the CsCl layer.) Cut off the tube bottoms (keep inverted) and resuspend the pellet in 100 µl 0.3 M NaOAc (pH 6) buffer. Transfer to a screw-cap 1.5 ml Eppi. Wash tube
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