Lecture 17

Question 1

stacking gels

Answer 1

the stacking gel is needed to concentrate/pack all the proteins in one band, so that they will start migrating in the running gel all at the same time. The running gel allows to separate the proteins in the sample based on their molecular weight

Question 2

relative mobility (Rf)

Answer 2

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

Question 3

Heating and running samples in a gel made with ~6 M ______ allows for proteins to denature but retain their native charge

Answer 3

urea; allows us to run a system with mobility affected by size AND charge

Question 4

Acid Urea Gels run ______ than SDS-PAGE and ________.

Answer 4

slower, backwards

Question 5

native (non-denaturing) gels

Answer 5

mobility of proteins depend on size, charge, and conformation; used to keep protein complexes intact

Question 6

2D protein electrophoresis

Answer 6

Proteins are separated based on 2 factors, in 2 dimension

Question 7

Dimension 1

Answer 7

isoelectric focusing gels; proteins will be separated by their overall pI (at their pI, they become uncharged and STOP)

Question 8

Dimension 2

Answer 8

SDS PAGE; resolved by size

Question 9

band identification

Answer 9

using antibody binding reactions or spot protein sequencing