Protein Purification Techniques

A technique for separating components in a mixture based on their density and size using centrifugal force.

A type of centrifugation where components are separated based on their sedimentation rate, which is influenced by their size and density.

A type of centrifugation where components are separated according to their density by layering them over a pre-formed density gradient.

A type of centrifugation where components are separated based on their density within a density gradient.

A technique for separating molecules based on their size by passing them through a selectively permeable membrane.

The movement of molecules from an area of high concentration to an area of low concentration.

The difference in concentration of a substance across a membrane.

A technique for separating proteins based on their charge and size.

A separation technique that separates molecules based on their size.

Smaller molecules get trapped inside the beads, while larger molecules move faster through the column.

The liquid used to carry the molecules through the chromatography column.

A type of chromatography used for separating proteins in a liquid environment.

A technique for analyzing and purifying proteins.

Different columns result in different levels of separation of proteins.

The peak in a chromatogram represents the different protein fractions separated by size.

Characterizing a protein involves identifying its properties, such as its size, shape, and function.

The process of detecting the presence of a protein.

The study of proteins.

A technique used to identify proteins in a sample. It involves separating proteins by size using electrophoresis, transferring them to a membrane, and then detecting specific proteins using antibodies.

A method used to determine the amino acid sequence of a peptide. Breaks down a peptide one amino acid at a time, starting from the N-terminus.

A technique that uses mass-to-charge ratio to identify and quantify molecules in a sample. Often used to determine the amino acid sequence of peptides and proteins.

The process of creating a new peptide sequence by chemically linking together amino acids.

A type of chemical synthesis that involves protecting the amine group of an amino acid with a 't-butoxycarbonyl' group (Boc) before linking it to another amino acid.

A chromatography technique that separates molecules based on their affinity to a specific ligand bound to a stationary phase.

In affinity chromatography, the molecule of interest binds to a specific ligand immobilized on a stationary phase.

In affinity chromatography, molecules that do not bind to the ligand are washed away in the first eluent.

To elute the molecule of interest from the stationary phase in affinity chromatography, a pH buffer is used to change the protein's charge.

Breaking down disulfide bonds and non-covalent interactions in affinity chromatography allows the molecule of interest to elute.

In ion exchange chromatography, molecules are separated based on their net charge.

The strength of interaction between a protein and an ion exchange matrix depends on the charge density of the protein.

The pH of the mobile phase in ion exchange chromatography can influence the charge of proteins.

A chromatogram in ion exchange chromatography visually represents the elution of different molecules.

In ion exchange chromatography, the elution profile of different molecules is affected by their charge and the strength of the interaction with the matrix.

A technique used to separate molecules based on their size and charge using an electric field.

A type of gel electrophoresis where proteins are denatured and coated with a negatively charged detergent, allowing separation based solely on size.

A type of gel electrophoresis that separates proteins based on their native charge and 3D shape.

A detergent used in SDS-PAGE to denature proteins and give them a uniform negative charge.

The isoelectric point of a protein is the pH at which it has no net charge and thus does not migrate in an electric field.

A technique used to separate proteins based on their isoelectric points by applying an electric field through a pH gradient.

A method that combines two electrophoresis techniques: isoelectric focusing and SDS-PAGE.

A type of stain used to visualize proteins in a gel electrophoresis experiment.

A type of stain used to visualize proteins in a gel electrophoresis experiment that is highly sensitive and can detect very small amounts of proteins.