Protein electrophoresis and SDS-PAGE

Question 1

what is protein electrophoresis?

Answer 1

it separates proteins based on size and charge
- uses electric field to move charged molecules through gel matrix

Question 2

what is sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)?

Answer 2

a specific type of electrophoresis that denatures proteins and coats them with negative charge, so they can be separated primarily by molecular weight instead of shape and change

Question 3

what is the native PAGE?

Answer 3

proteins are separated based on native conformation and charge

Question 4

What is the name for the abbreviation SDS-PAGE?

Answer 4

Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis

Question 5

what is the abbreviation for Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis?

Answer 5

SDS-PAGE

Question 6

What does the SDS-PAGE do?

Answer 6

denatures proteins with sodium dodecyl sulfate (SDS) and a reducing agent, then separating them using polyacrylamide gel electrophoresis

Question 7

What is the role of SDS?

Answer 7

An anionic detergent binding to proteins in a ratio proportional to their length, imparting a uniform negative charge
- denatures proteins, unfolding them into linear chains and masking their intrinsic charges, which results in them being separated by size

Question 8

What is the role of reducing agents?

Answer 8

Break disulfide bonds within and between protein molecules; ensure proteins are fully denatured and subunits of multi-subunit proteins are separated
- Examples of agents: 2-mercaptoethanol (BME) and diothiothreitol (DTT)