Biochemical Techniques and Instrumentation

Reagents

1. Subcellular fractionation buffer:

	Stocks	50 ml 1X solution
250 mM Sucrose	-	4.28 g
20 mM HEPES (7.4)	1 M	1 ml
10 mM KCl	-	0.0373 g
1.5 mM MgCl2	1 M	75 μl
1 mM EDTA	0.5 M	100 μl
1 mM EGTA	0.5 M	100 μl

HEPES : 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

EDTA – Ethylene diamine tetraacetic acid

EDTA : Ethylene glycol tetraacetic acid

At time of use: Take 10 ml and add the following:

	Stocks	50 ml 1X solution
1mM DTT	1M	10 μl
PI Cocktail (III)	-	50 μl

DDT: Dithiothreitol – a reducing agent

PI Cocktail (III)- Protease Inhibitor Cocktail