A protein was purified and we want to know the concentration of that protein in

Question

A protein was purified and we want to know the concentration of that protein in the final purification solution.

a. What property of the protein and specific amino acids responsible for that property will allow me to determine the concentration?

b. What amino acid contributes the most as an individual residue and by how much?

c. What is the name of the mathematical relationship between this property of the amino acids and the concentration of the protein?

d. What is the equation and what does each term stand for?

Explanation / Answer

a) The concentration of protein was calculated by complexing with metal ion like Cu+2 with amino group in amide group (C(=O)-N-H) of amno acids and aminoacids possesing groups like -CONH2,-CSNH2,-CH2NH2,-C(NH)NH2 i.e Cysteine,serine etc. this means compounds abosrb light energy at different wave length i.e absorption and estimated by colorimetry

b) aminoacids possesing groups like -CONH2,-CSNH2,-CH2NH2,-C(NH)NH2

c) A=ecl =C=A/Cl

In this construct calibration graph with known concentration of substrate and calcualte unknown.

d) A= absorption of protein or amino acid at particular wave length

e= molar absorbivity constant

l= length of tube

C= concentration of protein

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