☆ BSA ( s) -

make
10mg / ml BSA stock solution

Bradford (colorant)
%⑧qA→
*
weigh 1000mg
* Unknown sample

↓ addtelooml
LAB SKILL TEST
① water

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1-
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=

10mg / MIBSA

u
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↓ ↓ ↓ u

1:50 1:25 1 :
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part BSA
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part
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②⑧ ⑤ ☒
Getbigiomipipeuo.li 0.8 7.0
0.6
0mg 4mi mglml mglml
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mglml mglml BSA

BSA BSA BSA BSA

h h h h h
•
put 2✗ 20µL each diluted
stuck solution inte vessels Ng unknown
sample

③
Man Bk %% + Eeg
% % Bag BAM UM Umm MMMM
!° !°
M M 20Mt 20Mt
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20Mt
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20
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2×0-4 0 @
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Add 1mi
TOTAL : 76
14 vessels
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④
↓
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↓
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•

+
As blank for calibration

AND MIX ! Incubate for 10min after

↓ put in curettes
⑤
SPECTOPHOTOMETER
'

. I use blank limiisradpora) to calibrate spectrophotometer

2 measure every cuvette (0.2/0.4/0.610.817.08 unknown)
3 Repeat for every 2ⁿᵈ cuvette
⑥
BIOMEDICAL SCIENCE LABSKILLS TEST

DON’T WRITE ON THIS PAPER!!

Instructions:

• You need to determine the protein concentration in a “human urine”

sample as accurate and precise as possible using a standard
laboratory assay. During the test you need to fill in some questions
concerning the equipment that you used, the volumes that you used
etc. You need to generate a calibration curve that can be used to
determine the protein concentration in the test sample.
• When you are finished with the experimental work you need to show
the measurements that you’ve recorded to the supervisor. The
supervisor will sign-off on the results. After that you cannot change
the values anymore!
• Bring your laptop as you need it to make a calibration curve.
• Using the obtained data you need to plot the calibration curve in excel.
Finally, you need to report the protein concentration that you have
measured in the test sample. Be aware of significant figures when
reporting the final result.
• You are not allowed to talk to peers at any time during the test.
• You have exactly 1.5 hours to complete this test. When the end of the
exam is called, stop your activities and turn over your exam.
• Make sure you answered all the questions.
• Good luck!

Version 2023
Equipment and Materials
• Balance
• Spectrophotometer
• Glassware, pipettes and milli-q water, as required
• Plastic cuvettes, cuvette stirrers and tissues, as required
• 1 ‘urine sample’, being a normal water solution that should be treated as if it
were a urine sample. Write down the letter on the sample! Without this
information your work cannot be graded.
• Colour reagent for protein measurement
• Bovine Serum Albumin (BSA) (Sigma-Aldrich)
• Your personal laptop with Microsoft Excel or equivalent

Procedure
1. Prepare a stock solution of BSA of 10 mg/ml in milli-q water. Write down the
amount of BSA that you dissolved.
2. Prepare 5 dilutions using this stock solution to obtain a protein standard with a
range of 0.2 and 1.0 mg/ml BSA.
3. Pipet 20 µl each standard or unknown sample solution into separate disposable
cuvettes in duplicate. Write down the letter on the tube of the urine sample!
4. Add 1.0 ml of Bradford reagent to each cuvette and mix well.
5. Incubate at room temperature for 10 minutes.
6. Measure the absorbance of each sample at 595 nm.
7. KEEP THE CUVETTES UNTILL THE END OF THE TEST SO THAT THE SUPERVISOR
CAN COMPARE THEM TO YOUR RESULTS.

Analysis using Excel

1. Calculate the average absorbance for each sample of the calibration curve and
test sample.
2. Plot the absorbance as function of protein concentration (BSA).
3. Determine the protein concentration of the “human urine” sample using the
linear relationship obtained for the standard curve.
4. Show cuvettes and your results to the supervisor.

5. Clean-up workspace according to GLP guidelines.

Version 2023