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H, S, A, I, SG
He said all individuals SinG
Protein Separation Techniques (H) - answer✔Homogenization
disrupt cells, sonicator, centrifuge
Protein Separation Techniques (S) - answer✔Salting out
different proteins precipitate out at different salt concentrations
Protein Separation Techniques (A) - answer✔Affinity chromatography
add a tag; histidine binds; see how well there is an affinity for the group
Protein Separation Techniques (I) - answer✔ion exchange
separation based on charge; + proteins stick to - beads
Protein Separation Techniques (SG) - answer✔Size Exclusion/Gel Filtration
separation based on size
Cuts (protein separation techniques) - answer✔cut out what dont need/isnt protein
(use 40% concen. then keep liquid; if use 60% concen. then keep pellet)
Dialysis - answer✔protein separation based on bad with pores
1. Moles in bag + moles in buffer
2. Divide moles by total volume - new concentration
Electrophoresis - answer✔gels - move by size with electric current
1. Isoelectric focusing - purely pH
2. SDS page
pH separation is by pI and charge in 2d
Protein sequencing (who cuts after what) - answer✔Cyanogen bromide = methionine
trypsin = K, and R
chymotrypsin = F, M, L, W, Y
IP - answer✔Immunoprecipitation with proteins
1. add antibodies specific to protein of interest to lysed cells
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