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Michał Kaczmarek
equipment required:
spectrophotometer capable of measuring
absorbance in the 650 nm region.
disposable cuvettes for spectrophotometer
required reagents:
Reagent A – consist of 2% solution of Na2CO3 in 0,1 mol/dm3 NaOH
Regent B – consist of 0,5% solution of CuSO4 x 5H2O in 1% sodium citrate CH3COONa
Reagent C – to prepare reagent C mix both reagents (A and B) in appropriate molar ratio
5:1 (v/v)
protein standards BSA (bovine serum albumin) solutions at the concentration ranging from
0 µg/ml to 50 µg/ml
Folin-Ciocalteau reagent
Procedure Description:
The Lowry assay (1951) is an often-cited general use protein assay. Under alkaline conditions
the divalent copper ion forms a complex with peptide bonds in which it is reduced to a monovalent
ion. Monovalent copper ion and the radical groups of tyrosine, tryptophan, and cysteine react with
Folin reagent to produce an unstable product that becomes reduced to molybdenum/tungsten blue.
Procedure: