Native PAGE notes

Make up and run as with SDS-PAGE using the following buffers. Stain and destain as usual, too.

5 mL X% separating gel:

X/6 mL 30% acrylamide

1.25 mL 1.5 M Tris-HCl pH 8.8
3.75 - X/6 mL H2O
25 ?L 10% APS
2.5 ?L (5?L if X < 8) TEMED

5 mL 10% separating gel
1.67 mL 30% acrylamide
1.25 mL 1.5M Tris-HCl pH 8.8
2.08 mL H2O
25 ?L 10% APS
2.5 ?L TEMED

2 mL 5% stacking gel
1.15 mL H2O
335 ?L 30% acrylamide
500 ?L 0.5M Tris pH 6.8
15 ?L 10% APS
2.5 ?L TEMED

2x Native sample loading buffer
125 mM Tris HCl pH 6.8
0.05% bromophenol blue
20% glycerol

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Ornstein-Davis buffer formulations:

Resolving gel buffer: 1.5M Tris-HCl pH 8.8
Dissolve 27.23g Tris base in ~80ml deionized water
Adjust pH to 8.8 using HCl
Make up to 150ml with water

Stacking gel buffer: 1.0M Tris-HCl, pH 6.8
Dissolve 12g Tris base in ~60ml deionized water
Adjust to pH 6.8 using HCl
Make up to 100ml with water

Sample loading buffer: 62.5mM Tris, 10% glycerol, 0.025% bromophenol blue

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Alternative 5x Native sample loading buffer w/Ficoll PM-400
200 mM Tris HCl pH 8.8
0.05% bromophenol blue
10% w/v Ficoll PM-400

Native running buffer
190 mL 5x stock
810 mL nanopure H2O

5x stock running buffer
151 g Tris base
940 g Glycine
add H2O up to 9.5 L total volumep


Useful link here

OpenOffice spreadsheet of constituents here