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文献和实验Preparation of protein extracts for western blot 1.Grow cells to mid-log (OD600 less or equal to 1.0). 2.Harvest about 5 OD's of cells in a 13X100 glass dispo tube (Use new tubes.Can be up to about 6 mls of culture). 3.Pellet 3-5 min.in Tomy,3K
Use of Flow Cytometric Methods to Quantify Protein‐Protein Interactions
) 10‐ml Sephadex G25 gel filtration column (essentially any standard desalting column can be used) Amicon Ultra centrifugal concentrators (MWCO, 10,000; Millipore, cat
Preparation of protein extracts for western blot
buffer(or use 75μl of 3X Laemmli sample buffer). 9.Place in boiling water bath for 1 min. 10.Spin briefly in Tomy (at room temp--15 sec,3K),remove liquid with a ml pippetteman tip,and transfer to 1.5 ml microfuge tube.Spin in microfuge for 10 min. 11
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