REAGENT PREPARATIONBring all reagents to room temperature before use.Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mixgently until the crystals have completely dissolved. Add 20 mL of Wash Buffer Concentrate todeionized or distilled water to prepare 500 mL of Wash Buffer.Substrate Solution - Color Reagents A and B should be mixed together in equal volumeswithin 15 minutes of use. Protect from light. 100 μL of the resultant mixture is required per well.Calibrator Diluent RD5-26 (diluted 1:4) - Add 20 mL of Calibrator Diluent RD5-26Concentrate to 60 mL of deionized or distilled water to prepare 80 mL of Calibrator DiluentRD5-26 (diluted 1:4).