Dissociation medium

doi:10.1101/pdb.rec11497

Recipe

Dissociation medium

Reagent	Amount to add	Final concentration
1 M Na₂SO₄	20.44 mL	98 mM
0.5 M K₂SO₄	15 mL	30 mM
1 M MgCl₂	1.45 mL	5.8 mM
0.1 M CaCl₂	0.63 mL	0.25 mM
1 M HEPES (pH 7.4)	250 μL	1 mM
1 M glucose	5 mL	20 mM
Phenol Red (0.5%)	0.5 mL	0.001%
0.1 N NaOH	~0.1 mL	0.03 mN
H₂O	to 250 mL

Add above volumes to a sterile, dedicated 250-mL tissue culture bottle (pH should be ~7.4). Filter-sterilize using a 0.2-μm bottle filter. Store solutions at 4°C. (Na₂SO₄ and K₂SO₄ solutions should be stored at room temperature to avoid formation of precipitate.) Use dissociation medium within 2 wk.