Lecture 15-16

Question 1

Stacking gels

Answer 1

2 gels + 2 buffers + 3 pH’s = elegant solution

Question 2

top gel(stacking)=

Answer 2

5% acrylamide, pH 6.8 w/ Tris-HCl

Question 3

running gel=

Answer 3

~12% acrylamide, pH 8.8 w/ Tris-HCl

Question 4

upper buffer=

Answer 4

pH 8.3 w/glycine (pI = 6)

Question 5

acrylamide concentration effects

Answer 5

high% gel for small proteins

low % gel for large proteins.

Question 6

SDS-Page allows for separation of proteins

based on a single criteria􏰀–>

Answer 6

size

Question 7

Urea denaturing gels separate proteins based on

Answer 7

charge and size.

Question 8

NAtive denaturing gels separate proteins based on

Answer 8

charge, size and shape

Question 9

SDS page

Answer 9

– Proteins are boiled, denatured & coated w/ SDS
– Loaded into gel
– Small proteins = fast, Large proteins = slow
– MW size standards to determine protein length

Question 10

Gel staining – methods to visualize proteins

Answer 10

Coomassie blue – fastest, easiest, lowest sensitivity – Silver staining – time consuming, highest sensitivity – Fluorescent staining – mid-range

Question 11

2 dimension electrophoreses

Answer 11

size and isoelectric point

Question 12

ampholytes

Answer 12

molecules that can buffer at their pi

Question 13

First line of defence

Answer 13

• intact skin
• mucosal membranes
• normal microbiota

Question 14

second line of defence

Answer 14

• natural killer cells
• inflammation
• fever

Question 15

third line of defence (adaptive immunity)

Answer 15

t cells and b cells

-antibodies