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Lab Report Biochemistry (BIO 462)

The document is a lab report for an experiment determining protein concentration. It details that: 1) Coomassie Brilliant Blue dye was used in the Bradford assay and changes color from red-brown to blue when binding to proteins. 2) In the Lowry assay, peptides bind to copper ions. 3) A Bradford assay on a diluted sample found it contained 4.07 microL/mL protein originally, or 203.5 microL/mL in the original undiluted sample.

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Iman Farha
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186 views2 pages

Lab Report Biochemistry (BIO 462)

The document is a lab report for an experiment determining protein concentration. It details that: 1) Coomassie Brilliant Blue dye was used in the Bradford assay and changes color from red-brown to blue when binding to proteins. 2) In the Lowry assay, peptides bind to copper ions. 3) A Bradford assay on a diluted sample found it contained 4.07 microL/mL protein originally, or 203.5 microL/mL in the original undiluted sample.

Uploaded by

Iman Farha
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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LAB REPORT BIOCHEMISTRY

(BIO 462)

EXPERIMENT 2: PROTEIN DETERMINATION

NAME IMAN FARHA HANISAH BINTI DANIEL HASNI


STUDENT NUMBER 2019702307
GROUP AS2464B
LECTURER’S NAME DR NUR NADIAH MD YUSOF
DATE OF EXPERIMENT 13TH APRIL 2020
1. Name the dye used in Bradford assay

Coomassie Brilliant Blue

2. State the colour changes that occur when proteins combine with the dye reagent.

Red-brownish changes to blue

3. In Lowry protein assay, name the bond in protein that binds to copper ions.

Peptide bond

4. A Bradford assay was conducted to determine the total protein concentration in a sample.
A volume of 2 microL of the original sample was diluted to 100 microL with buffer before
performing the assay. The diluted sample gave an absorbance at 595 nm of 0.255. using
the data for the standards below,

Protein concentration (microL/mL) A595


25 0.008
125 0.087
250 0.113
500 0.197
750 0.295
1000 0.429
1500 0.608
a) Plot a standard curve based in the data given

Abs vs. protein concentration


0.7
0.6 0.608
y = 0.0952x - 0.1327
0.5 R² = 0.9439
0.429
Abs (595 nm)

0.4
0.3 0.295
0.2 0.197
0.1 0.113
0.087
0 0.008
25 125 250 500 750 1000 1500
-0.1
protein concentration (microL/mL)

b) Determine the concentration of protein the diluted sample


c) Calculate the total protein concentration in the original sample

b) Y=0.0952x-0.1327 c) M1V1=M2V2
0.255=0.0952x-0.1327 M1(2)=4.07(100)
X=4.07 microL/mL M1=203.5 microL/mL

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