UV Protein Determination.

Min Kuan 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
as followed:

		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.

Sincere,
Min.



	.
 




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