Jacob K. answered • 10/20/23
Experienced Tutor Specialized in Biochemical Sciences
%B refers to the percentage of a given buffer. When using a system like a chromatogram, often times two or more buffers are utilized. It does so, to allow for a efficient purification of a given substance. Even if only two buffers were used, a gradient can form, allowing elution to happen in steps, rather than all at once. This drives the several visualized peaks (and confusing looking graph).
To know the %B which elutes LDH from the column, you must know what peak contains your LDH. So, first, identify what peak has the highest activity of LDH. Then, look what time that peak occurred at during the grand scheme of the reaction. Lastly (depending on what program you are using), look what the buffer concentrations were at that point. Most programs have that information in a printout, or along with your graph in some way.
I will be up for a bit longer tonight in case you need emergent assistance and clarification. Feel free to message me to ensure that I see it. I am actively working with mLDH and hLDH in a lab right now, so it is fresh in my mind and I have an interest!
