Hi,

I am planning to run gradient elution for protein of my interest.
But I am stuck with gradient elution calculation.
I know some of the facts:
1. Buffer A shld be without salt
2. Buffer B shld have highest conc of salt (in my case 0.5 M)

I got following info from online sources:
A good rule of thumb is to use 10 times the column volume for your gradient. That means for a 10 ml column, a 100 ml gradient will work well. To prepare this kind of gradient, 50 ml of the buffer without salt is placed in the container that will plumb to the column and 50 ml of buffer with the final salt concentration is placed into the other container. A gradient concentration of 0 to 250-500 mM NaCl in the buffer will work well.

After reading this, I am confused with what numbers shld I plug-in in Gradient table in my method for IEC
Time Buffer A (%) Buffer B (%)

Please reply asap.
Your help is highly appreciated.