Dr. Wightman,
Here at U.C. Davis, Lactate Dehydrogenase was purified in one step
from beef steak (or heart muscle) using a NADH affinity column. The assay
was, of course, NADH to NAD+ monitoring absorbance at 340 nm. It took one
or two 5 hour lab periods. It was very simple and it taught them
purification using a Waring blender, affinity chromatography, fraction
collectors, spectrophotometer, and all about glycolysis. The experiment
was done in ice buckets and a cold room. Each lab had 30 students working
in pairs. (I am talking about the undergraduate biochemistry lab class,
formerly known as 101L. It has a new number, but I don't know it)
I was a T.A. years ago and they might have changed the lab by now,
but you can contact Dr. Larry Sprechman in Biochemistry and he can
provide you with all the information. We are also only an hour away from
you, so you come up here to see the set-up for yourself.
Good Luck, Jonathan Mazer, PhD candidate.
