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文献和实验Preparation of Competent Cells (from PMB)
#A 1X 200 ml 1X 10 mM MOPS pH 7.0 210 mg Free acid 230 mg Na salt 10 mM Rubidium chloride 240 mg Autoclave and store at 4 ℃ PMB #B 1X 200 ml 1X 10 mM MOPS pH 6.5 350 mg Free acid 85 mg Na salt 50 mM Calcium chloride 1.12 g 10 mM
[3H] Choline Labeling and TNF Treatment of HL-60 Cells
with ITS (4 ml/L should yield 5 mg/L insulin and 5 mg/L transferrin). --> "Appropriate" = bring cells to a final concentration of 2 X 105 cells/ml. 4) Add (0.5 µl/ml media) [3H] choline chloride to get ~0.5 µCi/ml. --> Specific activity of the [3H
Choline Labeling and TNF Treatment of HL-60 Cells
temperature PBS. 3) Resuspend final pellet in an appropriate volume of prewarmed serum free media and supplement with ITS (4 ml/L should yield 5 mg/L insulin and 5 mg/L transferrin). --> "Appropriate" = bring cells to a final concentration of 2 X 105
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