Lecture 13-14

Question 1

Assays should be…

Answer 1

Sensitive, specific, rapid, quantitative.

Question 2

Chromotography

Answer 2

Techniques to separate mixtures based on physical properties such as size or charge

Question 3

Stationary phase

Answer 3

a substance that the compounds to be separated pass

by or interact with

Question 4

mobile phase

Answer 4

the carrier for the compounds to be separated

Question 5

Size exclusion chromatography

Answer 5

Large molecules move the fastest, travel only in the Vo.

Question 6

Partition coefficient equation

Answer 6

KAV = (Ve – Vo)/ (Vt – Vo)

Question 7

Ion-exchange chromatography

Answer 7

Like charge or neutral charge will elute fastest.

Question 8

order of specificity of chromatography

Answer 8

1. Affinity chromatography
2. Ion exchange chromatography
3. Size exclusion chromatography

Question 9

Electrophoresis:

Answer 9

migration of ions in an electric field.Velocity is directly proportional to charge and inversely proportional to size and shape

Question 10

SDS

Answer 10

a detergent that will denature proteins and give them a negative charge.

Question 11

What is the gel composed of?

Answer 11

acrylamide, bisacrylamide, APS, TEMED, SDS and buffer

Question 12

Relative mobility( Rf)

Answer 12

how far your protein band goes compared to something fast(dye)

Question 13

Urea Denaturing gels

Answer 13

affected by size and charge/

Question 14

Native, non-denaturing gels.

Answer 14

Affected by size, charge and shape.

Question 15

partition coeffecient equation

Answer 15

KAV = (Ve – Vo) /(Vt – Vo)

Question 16

What is the partition coefficient of a large protein?

Answer 16

0

Question 17

What is the partition coefficient of a small protein?

Answer 17

1