Amino Acid Analysis and Protein Separation Techniques

A series of flashcards covering key concepts in amino acid analysis and various chromatography techniques for protein separation.

Amino Acid Analysis

The process of separating and detecting individual amino acids in a mixture.

Chromatography

A technique for separating components of a mixture based on different properties such as polarity or charge.

Thin Layer Chromatography (TLC)

A method where silica gel is spread as a thin layer on a sheet, and different components are separated as a solvent moves up the sheet.

Relative Mobility (RF)

The ratio of the distance moved by a sample to the distance moved by the solvent front during chromatography.

Ninhydrin

A reagent that reacts with amino acids to produce a purple color, used in detecting amino acids at low concentrations.

Fluorescamine

A reagent that reacts with amino acids to produce yellow fluorescence, used to detect amino acids when illuminated with UV light.

Ion Exchange Chromatography

A technique that separates molecules based on their charge, using resins that bind either cations or anions.

Gradient Elution

A technique in ion exchange chromatography where the concentration of salt is gradually increased to elute bound proteins.

Affinity Chromatography

A method that uses a ligand attached to the resin to selectively bind proteins with an affinity for that ligand.

SDS-PAGE

A type of gel electrophoresis that separates proteins based on size, using anionic detergent SDS that gives all proteins a uniform negative charge.

Isoelectric Focusing

A technique that separates proteins based on their isoelectric point, where the protein has no net charge.

Gel Filtration Chromatography

A method of separation based on molecular size, where larger molecules elute first from a gel matrix.

Enzyme Activity

The measure of the conversion of substrate to product per unit time by an enzyme, often used to assess the amount of enzyme present.

Specific Activity

The number of enzyme units per milligram of total protein, indicating the purity of an enzyme sample.

Mass Spectrometry (MALDI-TOF)

A technique for identifying proteins by measuring the time of flight of ionized particles based on mass-to-charge ratio.