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Aim: Principle:: To Quantify The Amount of Proteins Present in The Given Environmental Sample Using Biuret Test

To quantify protein concentration in a sample using the Biuret test. The test involves preparing standard solutions of bovine serum albumin (BSA) and measuring their absorbance with Biuret reagent. The unknown sample's absorbance is then used to estimate concentration by comparing to the standard curve. The protein concentration in samples x and y were found to be 2.47 mg/ml and 1.30 mg/ml respectively.

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gaurav saxena
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103 views2 pages

Aim: Principle:: To Quantify The Amount of Proteins Present in The Given Environmental Sample Using Biuret Test

To quantify protein concentration in a sample using the Biuret test. The test involves preparing standard solutions of bovine serum albumin (BSA) and measuring their absorbance with Biuret reagent. The unknown sample's absorbance is then used to estimate concentration by comparing to the standard curve. The protein concentration in samples x and y were found to be 2.47 mg/ml and 1.30 mg/ml respectively.

Uploaded by

gaurav saxena
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd

Aim: To quantify the amount of proteins present in the given environmental sample using Biuret test

Principle:

Requirement: Biuert reagent, standard stock solution of Bovine Serum Albumin (BSA)
of 10 mg/ml concentration, unknown sample, colorimeter & D/W.

Procedure: 1. Prepare a set of known standard varying concentrations of BSA from


0 to 5mg/ml with the interval of 1mg/ml, using the stock BSA and
distilled water given in table as given in the observation table.
2. In other test tubes labelled ‘x’ & ‘y’, pipette out 2ml of given protein
sample and add 3ml Biuret reagent (for ‘x’ and dilute it 1:1 with D/W
for ‘y’)
3. Incubate at room temperature for 10 minutes and then read against
blank at 540 nm.
4. Plot a graph of O.D (Y- axis) versus standard concentration of protein
(X-axis) and calculate concentration of protein in given sample by
extrapolating OD of sample onto X-axis.

Result: The protein concentration in the unknown sample ‘x’ & ‘y’ was found to
be 2.47mg/ml & 1.30mg/ml respectively.

Precaution: Magnesium and ammonium ions hinder the test. Use of excess of alkali
can, it can be removed.
Observation table:

Sr Required Amount of Distilled Biuret O.D at


Standard Stock BSA water (ml) Reagent 540nm
No.
concn(mg/ml) (ml) (ml)
1 0 0 2 3 0.00
2 1 0.2 1.8 3 0.07
3 2 0.4 1.6 3 0.15
4 3 0.6 1.4 3 0.23
5 4 0.8 1.2 3 0.33
6 5 1 1 3 0.38
7 x 2 0 3 0.19
8 y 1 1 3 0.10

Calculation:
For ‘x’: For ‘y’:
3mg/ml BSA  0.23 OD & ‘x’  0.19 OD 3mg/ml BSA  0.23 OD & ‘y’  0.10 OD
‘x’ = 2.47mg/ml ‘y’ = 1.30mg/ml
Concentration of ‘x’ by graph = 2.5mg/ml Concentration of ‘y’ by graph = 1.3mg/ml
& by calculation = 2.47mg/ml & by calculation = 1.3mg/ml

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