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Protein Concentration Analysis Methods

The document presents the results of protein determination using Biuret and Bradford assays, including absorbance data and calculated protein concentrations for various samples. For the Biuret assay, protein concentrations ranged from 343 µg/ml to 879 µg/ml, while for the Bradford assay, concentrations varied from 688 µg/ml to 749 µg/ml. The document includes tables and graphs illustrating the relationship between protein concentration and absorbance for both assays.

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91 views5 pages

Protein Concentration Analysis Methods

The document presents the results of protein determination using Biuret and Bradford assays, including absorbance data and calculated protein concentrations for various samples. For the Biuret assay, protein concentrations ranged from 343 µg/ml to 879 µg/ml, while for the Bradford assay, concentrations varied from 688 µg/ml to 749 µg/ml. The document includes tables and graphs illustrating the relationship between protein concentration and absorbance for both assays.

Uploaded by

alijhave
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

1

Name: Bilal Tahir Student ID: 0328555 Practical Session: 03

Principle of Life Sciences


Practical 2
Protein determination – Quantitative method

Biuret assay
Table 1: Protein concentration (µg/ml) and corresponding absorbances of each tube
Tube Protein
identification concentration(µg/ml) Absorbance
Blank 0 0.000
Standard 1 200 0.017
Standard 2 400 0.027
Standard 3 600 0.040
Standard 4 800 0.051
Standard 5 1000 0.066

2.5
y = 7E-05x
R² = 0.9936
2
Absorbance

1.5

0.5

0
0 200 400 600 800 1000 1200
Protein concentration(µg/ml)

Figure 1: Graph of Absorbance against Protein concentration(µg/ml) using Biuret


assay
2

Bradford assay
Table 2: Protein concentration (µg/ml) and corresponding absorbances of each tube
Tube Protein concentration
Identification (µg/ml) Absorbance
Blank 0 0.00
Standard 1 200 0.663
Standard 2 400 1.413
Standard 3 600 1.549
Standard 4 800 1.738
Standard 5 1000 1.792

2.5
y = 0.0022x
R² = 0.7819
2
Absorbance

1.5

0.5

0
0 200 400 600 800 1000 1200
Protein concentration(µg/ml)

Figure 2: Graph of Absorbance against Protein concentration(µg/ml) using


Bradford assay
3

Calculation of protein concentration of 3 samples for Biuret and Bradford


Assays

Biuret :

Using the trend line equation given in Figure 1, y = 7E-05x, we can find the
concentration of protein by substituting y with the Absorbance. Using this equation
for biuret samples

Sample 1: y = 7E-05x

0.024 = (7x10-5 ) x

X = 343 µg/ml

Sample 2: y = 7E-05x

0.0415 = (7x10-5 ) x

X = 593 µg/ml

Sample 3: y = 7E-05x

0.0615 = (7x10-5 ) x

X = 879 µg/ml
4

Bradford :

Using the trend line equation given in Figure 2, y= 0.0022x, we can find the
concentration of protein by substituting y with the Absorbance. Using this equation
for Bradford samples

Sample 1: y = 0.0022x

1.514 = 0.0022x

X = 688 µg/ml

Sample 2: y = 0.0022x

1.569 = 0.0022x

X = 713 µg/ml

Sample 3: y = 0.0022x

1.648 = 0.0022x

X = 749µg/ml
5

Table 3: Average absorbance of unknown samples with Biuret Assay and their
calculated protein concentration
Tube Biuret Assay Average Calculated protein
Identification Absorbance concentration(µg/ml)
Sample 1 0.0240 343
Sample 2 0.0415 593
Sample 3 0.0615 879

Table 4: Average absorbance of unknown samples with Bradford Assay and their
calculated protein concentration
Tube Bradford Assay Average Calculated protein
Identification Absorbance concentration (µg/ml)
Sample 1 1.514 688
Sample 2 1.569 713
Sample 3 1.648 749

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