10. Protein Analysis Flashcards
1
Q
A
2
Q
which moves first when separating based on size
A
larger elute SOONER
smaller elute later
3
Q
NATIVE Page
A
separate based on molecular radius and net charge
4
Q
SDS page
A
separate ONLY by molecular weight
- neg move to anode (+)
- small run faster than larger
5
Q
DTT/ 2-mercaptoethanol
vs
performic acid
A
both reducing agents that break disulfide bonds !
- performic (oxidizes DS.B)
- DTT etc (just breaks bonds)
6
Q
what is function of SDS
A
denature proteins and distribute a NEGATIVE charge to them
7
Q
what enzyme cleaves polypeptides into smaller fragments
A
endopeptidases
8
Q
what does say trypsin do for example
A
cleaves peptide bonds
9
Q
what is used to identify N terminal AAs
A
- FDNB / Sangers reagent
- Dansyl chloride (any primary amino group)
- aminopeptidase
10
Q
what is used to identify C terminal AAs
A
- carboxypeptidase
11
Q
edman degradation
A
removes N terminal AAs
