Lab Techniques in Molecular Biology

These flashcards summarize key concepts and terms related to laboratory techniques in molecular biology from the student's lecture notes.

Antibodies

Proteins made by the immune system that specifically bind to other proteins, used in various assays.

Western Blot

A laboratory method used to detect specific proteins in a sample through SDS-PAGE separation and antibody binding.

SDS-PAGE

A technique used to separate proteins based on their molecular weight.

Ponceau Red

A soluble dye used to stain proteins on membranes, allowing visualization of total protein banding patterns.

Primary Antibody

The antibody that binds specifically to the target protein in a western blot.

Secondary Antibody

An antibody that is raised against the primary antibody and is used for detection in assays.

HRP (Horseradish Peroxidase)

An enzyme conjugated to secondary antibodies that facilitates visualization of protein bands in western blots.

Enhanced Chemiluminescence (ECL)

A visualization method for western blots where luminol is oxidized by HRP to produce a light signal.

Affinity Chromatography

A method used to purify proteins by exploiting specific interactions between an affinity tag and its ligand.

Glutathione

A tripeptide that serves as a ligand for GST in affinity purification of proteins.

GST::EGFP Fusion Protein

A protein created by fusing glutathione-S-transferase with enhanced green fluorescent protein, used in purifications and assays.

Bradford Assay

A colorimetric protein quantification method that uses Coomassie G-250 dye.

Affinity Resin

Material used in affinity chromatography that contains a ligand for binding the target protein.

Column Capacity

The number of ligand molecules on an affinity resin available for binding target proteins.

Sonication

A physical method used for cell lysis involving high-frequency sound waves.

Freeze-Thawing

A physical lysis method where cells are frozen and thawed to disrupt cell membranes.

Coomassie G-250

A dye used in the Bradford assay that changes color upon binding to proteins.

Negative Control

A sample that is expected not to show any response or effect, used to validate assay results.

Molecular Weight

The mass of a molecule, often used to determine the size of proteins in SDS-PAGE results.

Protein Blocking

A step in the western blot process to prevent non-specific binding of antibodies to the membrane.

Affinity Tag

A specific sequence of amino acids used to facilitate the purification of proteins.