UV Protein Determination.
ckuan at kits.sfu.ca
Thu Sep 22 17:34:29 EST 1994
I am doing some UV protein scanning to esterminate protein
concentration of purified alpha-chymotrpsin, and crude alpha-chymotrpsin
that has Bovine Serum Albumin (BSA) in a ratio of 1:1 from
280nm to 340nm.
According to an assay method on this enzyme described in
Worthington Enzyme Manual (1972, p129), the calculation given is
Extinction coefficient: E(1%, 280) = 20.4
mg/ml = A(1cm,280nm) x 0.49
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ I use this underlined
formula to calculate the protein concentration for the purified protein,
right? The problem is, how do I find out the extinction coefficient
for the Crude Enzyme? I can't use the E for the purified enzyme (20.4)
since the crude one has BSA in it.
So, any ideas? :)
Thanks in advance.
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