Limitation of size exclusion chromatography
- Turbulence from layering sample over gel
- Diffusion during travel
- friction differences along the glass tube wall
What is the relationship between the charge of two molecule and their speed traveling down the column
same charge - moves fastest
neutral - slow
opposite charge - slowest
What amount of charge travels faster down the column
the greater the charge on a protein the slower it will travel
Affinity Chromotography
specific interaction between protein and trap (substrate|enzyme) that holds it in the column.
Order of Specificity
- Affinity chromatography
- Ion exchange chromatography
- Size exclusion chromatography
Electrophoresis
migration of ions in an electric field
Protein Gel Electrophoresis
Gel used to slow down movement and prevent diffusion of proteins and create separation.
SDS-PAGE
detergent that denatures proteins and give them a negative charge
Acrylamide
plastic monomer (toxic, carcinogenic, teratogenic), used industrially and can be chained together.
