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After washing several times in PBT (+ 0.1% BSA), staining was developed for 1 h in a solution (100 mM Tris Cl [pH 9.5], 100 mM NaCl, 5 mM MgCl 2 , 0.1% Tween 20, 1 mM Levamosole) containing 4-nitro blue tetrazolium chloride (0.23 mg/ml) and 5-bromo-4-chloro-3-indolyl-phosphatase (0.18 mg/ml) and then terminated in PBT containing 20 mM EDTA

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