Protein Separation and Identification Techniques

The study of all proteins produced by an organism, including their abundance, modifications, and interactions.

The process of determining which proteins are present in a sample.

Measuring the amount of each protein in a sample, including how quickly they're produced and degraded.

Determining the location of a protein within a cell or organism. This is crucial for understanding its function.

Changes made to a protein after it's been synthesized, impacting its function, stability, and interactions. Examples include phosphorylation and glycosylation.

The study of the functions of individual proteins, classes of proteins, and protein networks.

The study of the 3D structures of proteins, which is important for understanding their function and for drug development.

The study of how proteins interact with each other, including which proteins interact, when, and where.

A technique using a dye to visualize proteins on a gel, commonly used for its simplicity and affordability. It works by the dye binding to hydrophobic pockets in proteins.

A widely used method for identifying and detecting specific proteins in a sample. It involves separating proteins by size, transferring them to a membrane, and using antibodies to detect the target protein.

A crucial step in Western blotting where a blocking solution is used to prevent antibodies from non-specifically binding to the membrane.

A primary antibody binds specifically to the target protein in a Western Blot, identifying it.

A secondary antibody binds to the primary antibody, allowing for visualization of the target protein in Western blotting.

This method disrupts cell membranes by using chemical agents to dissolve lipids. Common detergents include Triton X-100 and SDS, making this approach effective for membrane proteins.

This procedure involves freezing and thawing cells repeatedly, causing ice crystals to form and disrupt cell membranes. It is a simple method but time-consuming and may not be efficient for all cell types.

This method uses organic solvents like ethanol or acetone to precipitate proteins, disrupting cell membranes. It is fast but may not be suitable for all proteins due to possible denaturation.

These agents, such as urea and guanidine hydrochloride, disrupt the hydrogen bonding network in proteins, aiding in solubilization. Effective for insoluble proteins but can also denature.

Enzymes like lysozymes are employed to break down bacterial cell walls, often in combination with other lysis methods for enhanced efficiency. A specific method that preserves protein integrity, but limited to bacterial cells.

This technique separates components based on density by spinning samples at high speeds. Heavier particles sediment at the bottom, leaving the lighter supernatant containing proteins.

This method involves altering the solubility of proteins, causing them to precipitate out of solution. This can be achieved using salts, organic solvents, or changes in pH.

This approach involves separating proteins based on their unique properties, such as size, charge, or affinity to specific ligands. Different methods are used to target specific properties.

A technique used to separate compounds by their different binding affinities to a specific molecule (ligand) immobilized on a stationary phase.

A process that separates proteins based on their net charge. It involves attaching proteins to charged resins and then eluting them by changing the buffer's ionic strength or pH.

A chromatography type where proteins are separated based on their size and shape. Smaller molecules move faster through the column, while larger molecules are slowed down.

A chromatography type that uses a stationary phase with a specific hydrophobic (water-repelling) surface. Hydrophobic proteins bind to the phase and are eluted using solvents that increase hydrophobicity.

A chromatography type used to separate based on specific interactions with a stationary phase. Separation occurs based on differences in the distribution of the components between the mobile and stationary phases.

A chromatography type where the stationary phase is composed of charged resins that interact with proteins based on differences in their net charge. Proteins bind to the stationary phase and are eluted by changing the ionic strength or pH of the buffer.

A chromatography type where a solid support matrix is used, and the proteins are separated based on their ability to bind to the matrix. It is a very versatile technique that can be used to purify proteins based on a wide variety of properties, such as size, charge, affinity, and hydrophobicity.

A method used to separate and purify proteins based on their size. It involves running proteins through a gel matrix, where smaller proteins migrate faster than larger proteins.

A technique used to separate proteins based on their size, using an electric current to move proteins through a gel matrix.

Polyacrylamide is a synthetic linear polymer that forms a gel-like matrix, acting as a sieve for separating proteins in electrophoresis.

SDS (sodium dodecyl sulfate) is a detergent that denatures proteins, giving them a uniform negative charge and allowing separation based solely on size.

A type of electrophoresis that separates proteins based on their native charge and size, without denaturing agents. It preserves the protein's native structure.

This technique combines two electrophoresis methods (IEF and SDS-PAGE) to separate proteins based on both their isoelectric point (pI) and molecular weight (MW).

This technique utilizes fluorescent dyes to compare protein samples on the same gel, highlighting differences in protein expression levels between samples.

This type of electrophoresis is used to study protein complexes (a group of proteins). It preserves the complex interactions between proteins.

A high-resolution technique that uses advanced software algorithms to analyze protein expression data from 2D gels, generating a 'map' that highlights differences in protein expression between samples.

A chromatography technique that separates proteins based on their size, with smaller proteins entering pores and eluting later than larger proteins that bypass the pores.

A chromatography method that separates proteins based on their hydrophobicity, with hydrophobic proteins binding to hydrophobic groups on a resin at high salt concentrations and eluting as the salt concentration decreases.

A method using semipermeable membranes and pressure to concentrate and desalt protein solutions, allowing small molecules to pass through while retaining proteins.

A technique using semipermeable membranes to separate molecules based on their size, allowing small molecules to diffuse out of a protein solution enclosed in the membrane.

The movement of charged molecules in an electric field, resulting in their separation based on their charge and size.

A technique that separates proteins based on their isoelectric point (pI), causing them to migrate in an electric field until they reach a pH where their net charge is zero.

A technique combining SDS-PAGE and IEF, providing high-resolution separation of complex protein mixtures.