In 2D-PAGE, what does the first dimension separate by, and what does the second separate by?
QUESTION
Identify the expected charge of a protein at pH<pI in an electric field.
QUESTION
What is the main role of SDS in SDS-PAGE sample preparation?
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ANSWER
Denatures proteins and imparts a uniform negative charge per mass. As an anionic detergent, SDS unfolds proteins and binds proportionally to length, providing consistent negative charge.