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Biochemical Tutorial Calculation QUESTIONS

The document provides instructions and questions for 6 problems involving calculations related to concentrations of biochemical compounds and proteins. It asks the reader to calculate volumes, masses, moles of compounds needed to achieve given concentrations, and the concentration of an unknown protein sample using absorbance data from a Bradford assay calibration curve.

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rejymol
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209 views

Biochemical Tutorial Calculation QUESTIONS

The document provides instructions and questions for 6 problems involving calculations related to concentrations of biochemical compounds and proteins. It asks the reader to calculate volumes, masses, moles of compounds needed to achieve given concentrations, and the concentration of an unknown protein sample using absorbance data from a Bradford assay calibration curve.

Uploaded by

rejymol
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
You are on page 1/ 9

Q1

What volume of a 7 mM solution of compound Z would you have to


add to 25 ml of water to make a 5 μM solution of Z?

03/03/2021 Biochemical Skills 1


Q2
Compound X has a molecular weight of 150 Da.
How much would you have to weigh out to make a 200 μM solution
in a total volume of 100 ml?

03/03/2021 Biochemical Skills 2


Q3
You are given 3 ml of a 50 mM solution of compound Y:

(a) How many μmol of Y are present in the sample?

(b) How many μmol of Y are present in 10 μl of the solution?

(c) If you wanted to remove a sample containing 75 nmol of Y, what


volume would you have to take?

03/03/2021 Biochemical Skills 3


Q4
Protein W has a molecular weight of 13 kDa. You have a purified
protein sample containing 10 ml of a 20 mg/ml solution. How many
μmoles of the protein do you have?

03/03/2021 Biochemical Skills 4


Q5
A typical bacterium has a diameter of 1 μm, and can be treated as
being a sphere. Calculate how many free protons are contained within
the bacterial cell at pH 7.

03/03/2021 Biochemical Skills 5


Q5
Respiratory metabolism is driven by the translocation of protons across the
cytoplasmic membrane into a region known as the periplasm. This
compartment has a volume that is 10 % of that of the entire cell. Assuming that
the pH here is also 7, how many free protons are there in the periplasm? How
many free protons are there in a single periplasm at pH 8?

03/03/2021 Biochemical Skills 6


Q6
The amount of protein within a solution can be determined colourimetrically using the
so-called Bradford Assay. This requires the construction of a calibration curve, using
known amounts of a standard protein (usually Bovine Serum Albumin, BSA) from
which unknowns can be estimated.
In a protein estimation by the Bradford assay, in which 1 ml of the Bradford reagent
was added to the protein sample + water (combined volume of 0.1 ml), the following
results were obtained for a standard curve:
Volume of 0.5 Dilution factor Absorbance at Concentration of BSA Concentration of
mg/ml BSA of BSA on 595 nm in mg/ml after BSA in mg/ml
added to making up to dilution with water ignoring 10 fold
assay (ml) 0.1 ml with dilution with water
water
0 0 0.92 0 0
0.01 0.1 1.06 0.05 0.5
0.02 0.2 1.21 0.10 1.0
0.04 0.4 1.50 0.20 2.0
0.06 0.6 1.60 0.30 3.0
0.08 0.8 1.65 0.40 4.0

0.01 and 0.03 ml samples of a protein solution of unknown concentration


gave, respectively, absorbance readings of 1.24 and 1.59 in the same assay
conditions.
03/03/2021 Biochemical Skills 7
Q 6 (a)
Plot a suitable calibration graph for protein estimation. Plot the data
by hand using graph paper.

03/03/2021 Biochemical Skills 8


Q 6 (b) & (c)
(b) What values do these readings give for the protein concentration in the
unknown solution (units should be in mg/ml)?

(c) Which value is likely to be the most accurate estimate of the unknown
protein concentration, and why?

03/03/2021 Biochemical Skills 9

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