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Leicester University Pathology Department Contracts REAGENT PRODUCTION METHOD Method Title: Annexin V Buffer Method number: 93 Version No: 01 Storage Temperature: 4ºC Pretreatment: Sterile Shelf Life: 6 months Molar Composition: 10mM HEPES 150mM NaCl 1mM MgCl2 5mM KCl 1.8mM CaCl2 Hazards: HEPES KCl CaCl2 Red star chemical Red star chemical Red star chemical Cross References: Chemicals and Equipment: Method: HEPES NaCl MgCl2 KCl CaCl2 = 5ml of 1M stock = 10mM final concentration = 15ml of 5M stock = 150mM final concentration = 0.5ml of 1M stock = 1mM final concentration = 2.5ml of 1M stock = 5mM final concentration = 0.9ml of 1M stock = 1.8mM final concentration (1.25ml = 2.5mM) Make up to approximately 450ml with UP H20 PH to 7.4 with NaOH Make up to 500ml with UP H20. Filter sterilise (under tissue culture hood) and store at 4ºC. Contract work\Masterdocumentstore\RPMs\RPM93