Table 2.

Detailed protocol for gel and electrophoresis experiments.

	Stacking gel	Resolving gel	10X Running Buffer	Light gel	Heavy gel	Moving solution	Conservation buffer	DTT solution	Iodoacetamine solution	Equilibration buffer	5X running buffer	Overlays solution
40% acrylamide/bisacrylamide (37.5:1) (w/v)	1 mL	6.3 mL
1.5M TRIS-HCl, pH 8.8		4.5 L		52.5 mL	52.5 mL	50 mL	100 mL			50 mL
0.5M TRIS-HCl, pH 6.8	2.5 mL
10% SDS (w/v)	0.1 mL	0.18 mL		2.1 mL	2.1 mL		4 mL
10% APS	0.05 mL	0.09 mL		2.1 mL	1.05 mL
TEMED	0.015 mL	0.009 mL		360 μL	60 μL
Glycine			120 g
SDS			40 g							20 g	40 g
TRIS			576 g								60.5 g
30% acrylamide/bisacrylamide (37.5:1) (w/v)				68 mL	128 mL
Glycerol					18 mL	74 g				378 g
1% bromophenol blue					210 μL							20 μL
1M DTT								0.1 g
Iodoacetamine									1.12 g
Equilibration buffer								10 mL	25 mL
Urea										355 g
Agarose type I-A												0.4 g
Agarose type VII												0.1 g
5X running buffer												20 mL
Apyrogenic water	6.35 mL	6.93 mL	to 4L	85 mL	14 mL	90 mL	296 mL			To 1L	to 4L	80 mL

Acrylamide/bisacrylamide (37.5:1) solution (w/v) (BioBasic Inc, Markham, Ontario, Canada), agarose type 1-A (Sigma, Oakville, Québec, Canada), ammonium persulfate (BioBasic Inc, Markham, Ontario, Canada), bromophenol blue (Sigma, Oakville, QC, Canada), dithiothreitol (DIT) (BioBasic Inc, Markham, Ontario, Canada), glycerol (BioBasic Inc, Markham, Ontario, Canada), glycine (BioBasic Inc, Markham, Ontario, Canada), iodoacetamide (BioBasic Inc, Markham, Ontario, Canada), sodium dodecyl sulfate (SDS) (BioBasic Inc, Markham, Ontario, Canada), TRIS (BioBasic Inc, Markham, Ontario, Canada), TRIS-HCI (BioBasic Inc, Markham, Ontario, Canada), tetramethylethylenediamine (TEMED) (BioBasic Inc, Markham, Ontario, Canada), urea (BioBasic Inc, Markham, Ontario, Canada), 2-hydroxythylagarose type VII (Sigma, Oakville, Quebec, Canada).