MB Lecture 7_Protein purification and SDS-PAGE OctNov 2024 _TZ

Protein purification

The process of isolating a specific protein from a mixture.

Cell lysis

The process of breaking down cell membranes to release cytosolic proteins.

Affinity chromatography

A technique used to separate proteins based on their binding affinity to a specific ligand.

6-Histidine tag (6-His)

A sequence of six histidine residues used to facilitate protein purification by metal ion affinity.

Ion Exchange Chromatography

A method that separates proteins based on their charge.

Gel filtration chromatography

A technique that separates proteins based on size as they pass through a porous gel.

SDS-PAGE

A method for separating proteins by size using polyacrylamide gel and the detergent sodium dodecyl sulfate.

Agarose beads

Solid supports used in affinity purification that can be conjugated to various ligands.

Imidazole

A compound used during protein elution in affinity chromatography to compete with histidine for binding.

Protein quantification

The determination of the concentration of protein in a sample.

Coomassie Brilliant Blue G-250

A dye used to stain proteins in polyacrylamide gels, allowing visualization.

Bovine serum albumin (BSA)

A commonly used standard protein for protein concentration assays.

Stacking gel

A low percentage acrylamide gel used in SDS-PAGE to concentrate proteins before they enter the resolving gel.

Resolving gel

The higher percentage acrylamide gel in SDS-PAGE where proteins are separated based on size.

Non-covalent interactions

Interactions that do not involve the formation of covalent bonds; important in dye binding to proteins.

pH gradient

A difference in pH across a system, significant in the movement of proteins during electrophoresis.