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CALCIUM-PHOSPHATE Transfection for Chromaffin Cells
Reagents and Solutions:
1. 2M CaCl2 - 11.1g CaCl2 (anhydrous) + 47.3 ml H2O (or 14.7g CaCl2-2H2O)
Filter sterilize and store at 4′C
2. 150 mM Na2HPO4 -2.13g Na2HPO4 + 99.7 ml H2O (or 4.02 g Na2HPO4-7H20 + 97.8 ml Store at 4′C (pH is >9, nothing will grow In it)
3. 2x PlBS
280 mM NaC1 1.84g
40 mM PIPES 1.21g
1.5 mM Na2HPO4 1.0 ml of 150 mM Na2HPO4
Dissolve components in approximately 80 ml H2O. AdJust pH to 6.95 with 1.0 M NaOH. Adjust volume to 100 ml. If desired, filter sterilize. Store at 4′C N.B.: The above amounts assume the free acid ot PIPES. For NaPIPES use 1.30g.