Composition of reaction mixtures
Weigh accurately 50mg of bovine serum albumin and dissolve
in distiiled water and make up to 50ml in a standard flask.
Working standard: Dilute 10ml of the stock solution to 50ml
with distilled water in a standard flask. One ml of this
solution contains 200μg protein.
500mg of the sample and grind well with a pestle and mortar
in 5-10 ml of the buffer.
and use the supernatant for protein estimation.
out 0.2,0.4,0.6, 0.8 and 1 ml of the working standard into a
series of test tubes.
out 0.1 ml and 0.2ml of the sample extract in two other test
up the volume to 1ml in all the test tubes. A tube with 1ml
of water serves as the blank.
5ml of reagent C to each tube including the blank. Mix well
and allow to stand for 10min.
add 0.5ml of Folin-Ciocalteau reagent, mix well and incubate at
room temp in the dark for 30 min. Blue colour is developed.
the readings at 660nm.
a standard graph and calculate the amount of protein in the
sample. Express the amount of protein in mg/g or 100g sample.
LOWRY OH, ROSEBROUGH NJ,
FARR AL, RANDALL RJ. (1951) Protein measurement with the Folin
phenol reagent. J Biol Chem. 193(1):265-75 (Click
here to download).