Bio-Rad BioLogic LP System - Manual (Page 17)

In Manual mode:
a. Press the PUMP soft key, then press FLOW.
b. Set flow rate to 1.5 ml/minute, press OK. Press START.
c. Press the UV soft key, press SET RANGE.
d. Using the INCREASE and DECREASE soft keys, set UV monitor
range to 0.05 AUFS.
e. Press OK.
f. Press ZERO—the system will set the UV reading to zero.
g. Press the PUMP soft key.
h. Press the STOP soft key.
Section 18
Programming the Pump Method
The BioLogic LP starter kit allows you to separate a standard mixture
containing equine myoglobin, conalbumin, chicken ovalbumin, and soybean
trypsin inhibitor, using a Bio-Scale Mini Macro-Prep High Q cartridge.
Buffer “A” is 25 mM Tris HCl, pH 8.1; buffer “B” is buffer “A” with 0.5 M
NaCl. Equine myoglobin is not retained on the column—it elutes in the
void volume (first peak). The remaining three proteins bind to the column,
and elute separately as the salt concentration increases.
1. Press the PROGRAM soft key, select NEW METHOD.
2. Select Time programming mode.
3. Enter the pump program.
4. Press ADD.
5. Select buffer “A” with the Previous/Next keys, press OK.
6. Enter step length of 3 minutes, press OK.
7. Enter flow rate of 1.5 ml/minute, press OK.
8. You have now entered the first step of the method (Buffer “A”, step
length 3 minutes, flow rate 1.5 ml/minute).
9. Enter the remaining steps:
Step 2. Gradient 0% to 50% “B”, step length 10 minutes, flow rate
1.5 ml/minute.
Step 3. Buffer “B”, step length 6 minutes, flow rate 1.5 ml/minute.
Step 4. Buffer “A”, step length 6 minutes, flow rate 1.5 ml/minute.
10. After entering Step 4, press OK.
13
4006073D.qxp 8/17/2009 7:45 AM Page 13