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Revision as of 00:23, 3 October 2012 (view source) Acker (Talk | contribs) (→SDS PAGE Gel Electrophoresis) ← Older edit		Revision as of 00:24, 3 October 2012 (view source) Acker (Talk | contribs) (→SDS PAGE Gel Buffer Information) Newer edit →
Line 16:		Line 16:
	2.0 ml 1M Tris-HCl pH 6.8		2.0 ml 1M Tris-HCl pH 6.8
		+
	0.8 g SDS		0.8 g SDS
		+
	4.0 ml 100% glycerol		4.0 ml 100% glycerol
		+
	0.4 ml 14.7 M β-mercaptoethanol		0.4 ml 14.7 M β-mercaptoethanol
		+
	1.0 ml 0.5 M EDTA		1.0 ml 0.5 M EDTA
		+
	8 mg bromophenol Blue		8 mg bromophenol Blue
Line 25:		Line 30:
	288 g glycine		288 g glycine
		+
	60.4 g Tris base		60.4 g Tris base
		+
	20 g SDS		20 g SDS
		+
	1.8 L H2O		1.8 L H2O

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Revision as of 00:24, 3 October 2012

SDS PAGE Gel Electrophoresis

General Procedure

1. Place gel cartridge in gel box with loading buffer
2. Load samples
3. Run the gel
4. Place gel in DI water for one hour
5. Remove water, add staining agent (GelCode Blue), leave on rocker overnight
6. Remove GelCode Blue, add DI water. Refresh DI water every hour until gel bands are visible

SDS PAGE Gel Buffer Information

4X Loading Buffer

2.0 ml 1M Tris-HCl pH 6.8

0.8 g SDS

4.0 ml 100% glycerol

0.4 ml 14.7 M β-mercaptoethanol

1.0 ml 0.5 M EDTA

8 mg bromophenol Blue

10X Running Buffer

288 g glycine

60.4 g Tris base

20 g SDS

1.8 L H2O

1. Take protein sample, add Loading Buffer. Boil at 95 degrees Celsius for ten minutes
2. Remove premade gel cartridge from package, remove comb, rinse wells with DI water
3. Insert cartridge into gel box, add running buffer to top and bottom wells
4. Add samples into separate wells, along with a protein ladder.
5. Run gel at 200V for around one hour