AG Daniel:Protocols

AG Daniel

protein analysis

Purification:

protocol	recipe
x harvest cells
x wash with PBS
x lyse or freeze at -80°C	50mM	Tris (pH 7,5)	0,33g
	270mM	Saccharose	4,62g
	1%	Triton X100	0,5 ml
	ad	dd H2O	50 ml
	1 pill	Complete Protease Inhibitor
	aliquot and store at -20°C
x incubate on ice for 30min
x spin at max speed for 15min at 4°C
x transfer supernatant (=protein) in new tube, store at -20°C
x determine protein amount with BCA method
x prepare protein for loading (with 5x loading buffer)	300mM	Tris (pH 6,8)	0,47g
	50%	Glycerol	5ml
	2%	SDS	0,2g
	3%	β-MercaptoEtOH	0,3ml
	1 Sp	Bromphenolblue
	ad	dd H2O	10ml

SDS-PAGE:
Western Blot:
Detection: