Lecture 15

Question 1

Large proteins travel in only…

Answer 1

the Vo (equals Ve)

Question 2

moderate sized proteins travel in…

Answer 2

Vo and some Vi

Question 3

small proteins travel in..

Answer 3

only Vt (equals Ve)

Question 4

partition coefficient

Answer 4

the fraction of volume of the column available to sample

Question 5

Kav=

Answer 5

(Ve-Vo)/(Vt-Vo)

Question 6

RMW

Answer 6

relative molecular weight; the shape of the protein of interest may be different from the size standards causing it to run faster/slower than the actual molecular weight

Question 7

If Kav is ____ or ___ you will get a poor separation of proteins

Answer 7

> 0.8 or <0.2

Question 8

Samples become diluted as:

Answer 8

• turbulence from layering sample over gel
• diffusion during travel
• friction differences along the glass tube wall

Question 9

It is probably optimistic to expect the sample to become diluted..

Answer 9

3x

Question 10

Ion Exchange Chromatography

Answer 10

proteins in a sole get carried down by gravity and will get slowed by beads based on charge; like-charges or neutral elute quickly

Question 11

anion exchange

Answer 11

column is + and targets are -

Question 12

cation exchange

Answer 12

column is - and targets are +

Question 13

In Ion Exchange, altering pH reduces…

Answer 13

charge of proteins bound to column

Question 14

In Ion Exchange, increasing salt conc…

Answer 14

increase competition for binding the stationary phase

Question 15

Ionexchangecolumnsgiveafast/clean method to extract DNA from….

Answer 15

cleared bacterial lysate