Unit 4.1 Flashcards
1
Q
Why is protein purification required?
A
to study structure and function
2
Q
What challenges come with purifying proteins?
A
- the mixture of proteins in cells/tissue
- diversity of proteins (such as size, charge, binding characteristics, and cellular location)
3
Q
what determines a protein’s separation/purification parameters and assay?
A
It’s size, charge, binding characteristics and cellular location
4
Q
What is the order of the protein purification workflow?
A
- identify source (blood, cell type, etc)
- fractionation/enrichment (by centrifugation or salt fractionation)
- Purification
- monitoring purification (gel electrophoresis)
- quantification (protein concentration or enzyme activity)
- analysis (structure and function)
5
Q
how does gel electrophoresis work?
A
molecules are separated in electrical field through a porous field acting as a molecular sieve.
6
Q
what medium is usually used?
A
Polyacrylamide (proteins) and sometimes Agarose for nucleic acids
7
Q
What type of proteins is electrophoresis used for?
A
both native and denatured, but mostly denatured