Measuring protein activity

Question 1

Why measure proteins?

Answer 1

Identity
Concentration
Activity

Question 2

What is spectroscopy?

Answer 2

The study of how electromagnetic radiation interacts with matter

Question 3

Energy carried by electromagnetic radiation is dependent on what?

Answer 3

Wavelength

Frequency

Question 4

How do we work out energy in spectroscopy?

Answer 4

E=hc/lambda

Question 5

Name the three types of spectroscopy

Answer 5

NMR
Infrared
Ultraviolet

Question 6

What is wavelength dependent on?

Answer 6

Structure

Concentration

Question 7

What wavelength do aromatic residues absorb?

Answer 7

280nm

Question 8

What is a peak at 200-230 caused by?

Answer 8

Peptide bonds

Carboxylic moieties

Question 9

Explain the basic set up of a spectrophotometer

Answer 9

Light source
Monochromator
Cuvette (containing sample)
Output

Question 10

Absorbance is dependent on three things. What are they?

Answer 10

Concentration of solute
Light path length
Molar absorption/ extinction coefficient

Question 11

What is the Beer-Lambert relationship?

Answer 11

Alambda = epsilonlambda x c x l

Question 12

Absorbance is proportional to ________

Answer 12

Concentration

Question 13

What is stoichiometry?

Answer 13

Remembering the ratios involved in couple reactions
if 1 A = 2 Bs
and 1 B = 2 Cs
you must remember 1 A is 4 Cs