Bio-Rad Gel Doc™ EZ System - Manual (Page 33)

SDS-PAGE
19
Criterion Tris-HCI Gel Separation Guide
Tris-HCl gels are available in a wide selection of single percentages and
gradients for the separation of proteins by SDS-PAGE.
SDS-PAGE Buffers
Sample Preparation
Determine the appropriate protein concentration of your sample based on the
detection method and load volume used. Dilute 1 part sample with 1 part
sample buffer and heat at 95ºC for 5 min.
Running Conditions
Tris-HCI Gels Optimal Separation
10% 30-150 kD
4-20% gradient 10-200 kD
Running buffer, 1x 25 mM tris
192 mM glycine
0.1 SDS
DO NOT ADJUST pH
Sample buffer 62.5 mM Tris-HCI, pH 6.8
2% SDS
25% glycerol
0.01% Bromophenol Blue
5% 2-mercaptoethanol or 350 mM DTT (added fresh)
Power conditions 200 V constant
Starting current 90-120 mA/gel
Final current 35-55 mA/gel
Run time 55 minutes