SDSpolyacrylamide gel electrophoresis

SOLUTIONS

30%acrylamidemix	29%(w/v)acrylamide
	1%(w/v)N,N""-methyleNEBisacrylamide
5xElectrophoresisbuffer	Trisbase:15.1g
	Glycice:94g
	SDS:5g
	H2Oupto1liter
4xResolving-gelbuffer	1.5MTris-Cl(pH8.8)
8xStacking-gelbuffer	1MTris-Cl(pH6.8)
1xSDSgel-loADIngbuffer	50mMTris-Cl(pH6.8)
	100mMdithiothreitol
	2%SDS
	0.1%bromophenolblue
	10%glycerol

RESOLVINGGELMIXTURE

ResolvingGel(30ml)	6%	8%	10%	12%	15%
H2O	16.2	14.2	12.2	10.2	7.2
30%acrylamidemix	6.0	8.0	10	12	15
4xresolving-gelbuffer	7.5	7.5	7.5	7.5	7.5
20%SDS	0.15	0.15	0.15	0.15	0.15
20%APS	0.15	0.15	0.15	0.15	0.15
TEMED	0.024	0.018	0.012	0.012	0.012

ResolvingGel(10ml)	6%	8%	10%	12%	15%
H2O	5.4	4.7	4.1	3.4	2.4
30%acrylamidemix	2.0	2.7	3.3	4	5
4xresolving-gelbuffer	2.5	2.5	2.5	2.5	2.5
20%SDS	0.05	0.05	0.05	0.05	0.05
20%APS	0.05	0.05	0.05	0.05	0.05
TEMED	0.008	0.006	0.004	0.004	0.004

STACKINGGELMIXTURE

StackingGel(10ml)
	H2O	6.9ml
	30%acrylamidemix	1.7ml
	8xstacking-gelbuffer	1.25ml
	20%SDS	0.05ml
	20%APS	0.05ml
	TEMED	0.01ml

SOLUTIONSFORSTAININGANDDESTAINING

Stainingsolution	methanol	90ml
	H2O	90ml
	aceticacid	20ml
	CoomassieBrilliantBlueR250	0.25g
	Dissolvewellandfiltratethroughafilterpaper.
Destainingsolution	isopropanol	12.5%
	glacialaceticacid	10%
	AdditionofsmallpiecesofusedX-rayfilmwiththegelwillacceleratethedestainingprocess.

SOLUTIONFORDRYING

20%methanol,3%glycerolindiltilledwater