GE Amersham ECl Gel Box User Manual page 42

5 Protocols
5.1 Electrophoresis protocol
Sample loading
Running conditions
42
To load the samples, follow the instructions:
Step Action
1 Prepare the samples by adding sample and 2× sample buffer in
a 1:1 mixture.
Example: for 10 μl sample, add 10 μl 2× sample buffer.
Note:
For native conditions: use a sample buffer without SDS and DTT.
2 Heat the samples at 95°C for 5 minutes.
3 Spin down the samples quickly in a microcentrifuge and load the
samples directly into the wells in the gel.
Note:
A maximum of 0.5 μg/band per well of sample can be loaded.
Overloading may cause smearing and distortion.
Note:
To ensure uniform mobility, load an equal volume of 1× SDS
sample buffer into any unused well.
4 Place the safety lid on top of Amersham ECL Gel Box.
Run the gel according to the recommendations below.
Parameter
Voltage
Expected current
Value
160 V
Start: approx. 45 mA
End: 23 to 33 mA
Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC