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文献和实验from the cultures. Place the sample in a separate test tube (it need not be sterile). 2. Dilute 4 parts of stock Trypan Blue with 1 part of 5X saline and add 0.1 ml of the diluted dye to your sample. Mix gently. 3. Set up a hemocytometer and cover slip
you need: Sample Buffer: 0.06M Tris-HCl, pH 6.8 10% (v/v) glycerol 2% (w/v) SDS 5% (v/v) 2-mercaptoethanol 0.0025% (w/v) bromophenol blue Grow cells overnight (~1E7 cells/ml) and collect 1.5 ml cells in 1.5 ml microfuge tube (1 minute, 14000xg).
In Situ Cell Death (Apoptosis) Detection by TUNEL labeling
% EtOH, 2 min. dH2O rinse.Incubate slides in a 1µg/ml Proteinase K/10mM Tris solution, 15 min., RT. (7.5µl of 20µg/µl PK in 150 ml 10mM Tris, pH 7.4-8.0).All slides: 1x PBS rinse, 2 times (+ 10 min for those non-positive control slides).(Positive
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