Protein structure of a helix, B sheet, reverse turn - answer a: LEMARKHQ
B: VICYFTW
RT: PSDNG
how many amino acids in alpha turn helix - answer3.6
Super secondary structure - answerhelix turn helix
parallel vs. antiparallel - answerparallel wedges face same way
antiparallel face opposite
Isoelectric point (pI) - answerpH at which a particular molecule carries no net electrical
charge.
Protein Separation Techniques (ALL) - answerHomogenization
Salting Out
Affinity Chromatgraphy
Ion Exchange
Size Exclusion/Gel Filtration
H, S, A, I, SG
He said all individuals SinG
Protein Separation Techniques (H) - answerHomogenization
disrupt cells, sonicator, centrifuge
Protein Separation Techniques (S) - answerSalting out
different proteins precipitate out at different salt concentrations
Protein Separation Techniques (A) - answerAffinity chromatography
add a tag; histidine binds; see how well there is an affinity for the group
, Protein Separation Techniques (I) - answerion exchange
separation based on charge; + proteins stick to - beads
Protein Separation Techniques (SG) - answerSize Exclusion/Gel Filtration
separation based on size
Cuts (protein separation techniques) - answercut out what dont need/isnt protein
(use 40% concen. then keep liquid; if use 60% concen. then keep pellet)
Dialysis - answerprotein separation based on bad with pores
1. Moles in bag + moles in buffer
2. Divide moles by total volume - new concentration
Electrophoresis - answergels - 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) - answerCyanogen bromide = methionine
trypsin = K, and R
chymotrypsin = F, M, L, W, Y
IP - answerImmunoprecipitation with proteins
1. add antibodies specific to protein of interest to lysed cells
2. add antibody binding protein on beads
3. centrifuge and protein is on bead with antibodies
4. gel elect
CO-IP - answerobserve protein protein interaction
Messelson Stalh - answerRATSHIT.
Western Blotting - answerrun gel - transfer to sheet - stained with radioactive antibody -
autoradiogram - exposed antibody
ELISA - indirect and sandwich - answerEnzyme linked immunosorbant assay
1.) indirect - antigen coated well - bind persons antibody - bind antigen with enzyme -
bind substrate = rate of color
2.) Sandwich - antibody coated well - bind person antigen - bind 2nd antibody with
enzyme - bind substrate = rate of color
ESI - answerelectric spray ionization - charged molecules - time of flight detector
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