Gel strength
Preparation of 12% (w/v) Polyacrylamide SDS Gel should Polymerize in 20-30 minutes
Electrophoresis test
Electrophoresis of Protein samples and Unstained Protein Molecular Weight Marker in a 12% Tris-glycine SDS gel should resolve individual clear bands
Visualization of bands after staining
Crisp Blue Bands should be visualized in a clear background after staining with Coomasie Blue Stain and destaining
Description
SDS PAGE technique involves separation of proteins based on their size under denaturing conditions using sodium dodecyl sulphate (SDS) and beta mercaptoethanol (BME). SDS cleaves non ionic interactions while BME cleaves disulphide bonds. SDS also imparts a strong negative charge to the polypeptides thereby their charge to mass ratio is rendered uniform. This allows the polypeptides to be separated on the basis of only their size, which is read by comparison with a standard marker.
Includes
Bis-Acrylamide solution - 55 ml
4X Separating buffer - 35 ml
4X Stacking buffer - 10 ml
10% SDS - 2 ml
Distilled water -100 ml
Ammonium Per Sulphate(APS) - 2 vials
TEMED - 120 mcl
Mid range marker 1 - 350 mcl
Unknown Protein samples - 5 Nos.
10X Tank Buffer Stock - 200 ml
Coomassie Brilliant Blue stain - 250 ml
Destain Solution I - 250 ml
Destain Solution II - 250 ml
