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发表于 2014-6-10 21:02:01
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看图形钙离子没有被刺激出来，会不会实验步骤中还需要孵育？或者孵育用的缓冲液？
见下面1990年经典文章中的步骤：
PBL (10E7cells/ml) were incubated at 37°C for 20 min in HBSS containing fluo-3/AM in a final concentration of 4uM. Next, PBL were diluted 1/5 in HBSS containing 1% fetal calf serum and incubated for 40min at 37 ℃. The ceils were then washed three times and resuspended at a concentration of 1 x
10E6cells/mi in a Hepes-buffered saline, containing 137 mM NaCl, 5 mM KCl, 1 mM Na2HP4, 5mM glucose, 1 mM CaCl2, 0.5 mM MgCl2, bovine serum albumin 1 g/l, Hepes 10 mM, pH 7.4.
Before each assay, the cells were incubated for 10 min in a 37°C water bath. The cells were subsequently stimulated with mAbs and lectins at the indicated concentrations and analysed on a Facscan flow cytometer
全文PDF见：
PBL_fluo3-am.pdf
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发表于 2014-6-10 17:12:28
