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文献和实验a change in specific activity. 3.2. Preparation of Wnt3A CM for Fractionation 1. To the filtered CM from Section 3.1 , add Triton X-100 to1% (v/v), Tris-HCl, pH 7.5, to 20 mM, and NaN 3 to 0.01%(w/v) (e.g., to 930 mL CM, add 50 mL of 20% (v/v) TritonX
(optional; see Steps 21 and 37) Horizontal gel electrophoresis tank for agarose gels (for duplex PCR only; see Steps 40-53) Standard DNA sequencing gel apparatus with 31 x 38.5-cm glass plates, 0.4-mm spacers, and a shark's-tooth comb (for hot-stop PCR
setting) and collect the mononuclear cell (MNC) (buffy coat) layer. Resuspend and wash MNCs x 3 with Dulbecco's Phosphate Buffered Saline (DPBS) with 10% Fetal Bovine Serum (FBS) x 10 min, 515 g, before plating into two 12-well plates in full EPC growth
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