, Youssef Farhat, MD/PhD Pathology Resident Medical College of Wisconsin. pNNP is sensitive to light and thus should be protected. A: HEPES (C 8 H 18 N 2 O 4 S MW: 238.3 g/mol) B: Distilled water; To prepare L of HEPES Buffer ( M, pH 6.8~8.2 ) Input buffer volume, molar concentration to get formula. pH should be 7.2 to 7.8, conductivity should be 14,000 to 16,000. Adjust pH to 7.4 with NaOH. © ELISA Genie. Required components. Sterilize by filtration through a 0.45-μm nitrocellulose filter. Add one NaOH pellet to raise the pH towards 7.4. Alkaline phosphatase catalyzes the hydrolysis of pNPP to pNP. Begin monitoring pH of the solution.
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