Henderson-Hasselbach Equation - ANSWER pH = pKa + log ([A-] / [HA])
FMOC Chemical Synthesis - ANSWER Used in synthesis of a growing amino acid chainto a polystyrene bead. FMOC is used as a protecting group on
the N-terminus.
Salting Out (Purification) - ANSWER Changes soluble protein to solid precipitate. Protein precipitates when the charges on the protein match the
charges in the solution.
Size-Exclusion Chromatography - ANSWER Separates sample based on size with smaller molecules eluting later.
Ion-Exchange Chromatography - ANSWER Separates sample based on charge. CM attracts +, DEAE attracts -. May have repulsion effect on like
charges. Salt or acid used to remove stuck proteins.
Hydrophobic/Reverse Phase Chromatography - ANSWER Beads are coated with a carbon chain. Hydrophobic proteins stick better. Elute with non-Hbonding solvent (acetonitrile).
Affinity Chromatography - ANSWER Attach a ligand that binds a protein to a bead. Elutewith harsh chemicals or similar ligand.
SDS-PAGE - ANSWER Uses SDS. Gel is made from cross-linked polyacrylamide. Separates based off of mass with smaller molecules moving
faster. Visualized with Coomassie blue.
SDS - ANSWER Sodium dodecyl sulfate. Unfolds proteins and gives them uniform negative charge.
Isoelectric Focusing - ANSWER Variation of gel electrophoresis where protein chargematters. Involves electrodes and pH gradient. Proteinstops at
their pI when neutral.
FDNB (1-fluoro-2,3-dinitrobenzene) - ANSWER FDNB reacts with the N-terminus of the protein to produce a 2,4-dinitrophenol derivative that labels
the first residue. Can repeat hydrolysis to determine sequential amino acids.
DTT(dithiothreitol) - ANSWER Reduces disulfide bonds.
Iodoacetate - ANSWER Adds carboxymethyl group on free -SH groups. Blocks disulfide bonding
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