Laboratory: Experiment 2: SDS PAGE

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Laboratory Experiment 2 SDS PAGE Lecture
SDS Polyacrylamide Gel Electrophoresis In-Class
Writing review proposals (pages xxxiii-xxxvi
129-133) Read Unit 2 (34-35) JBC 266 3811-14
(1991) Hand In proposal (3 copies) Due Next
Class flow chart 4 (day 1 DNA extract)
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SDS- Polyacrylamide Gel Electrophoresis
2-mercaptoethanol reduces S-S bonds SDS
denatures proteins imparts uniform
negative charge SDS-PAGE separates most proteins
by size. Some proteins migrate faster or slower
than expected.
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stacking gel 4 acrylamide, pH 6.8
resolving gel 15 acrylamide, pH 8.8

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His-tagged FnbA protein 138 amino acids MW
15,341
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MRGSHHHHHHGSGGQNSGNQSFEEDTEEDKPKYGQGGNIVDIDFDSVPQI
HGQNKGNQSFEEDTEKDKPKYEHGGNIIDIDFDSVPHIHGFNKHITEIIE
EDTNKDKPSTQFGGHNSVDFEEDTLPKVSSGLQPSLISstop M
start HHHHHH 6 His tag RGSGSG SGLLIS
vector GQN KVS FnbA
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