Protein Separation

Sodium Dodecyl Sulphate Polyacrylamide

Meaning of SDS PAGE

Agarose

Gel used in gel electrophoresis of nucleic acids

Polyacrylamide gel

Gel used in gel electrophoresis for proteins

Native PAGE and SDS PAGE

Two types of PAGE used in proteins

Native PAGE

It is the type of analysis of proteins by non-denaturing electrophoresis

Tris-electrophoresis

Buffer used in Native PAGE

SDS PAGE

The most widely used method for characterization of proteins and evaluation of their molecular weights; as well as for establishing homogeneity of protein samples

Tertiary and quaternary

What two protein structures must be distributed completely to achieve protein separation according to molecular weight?

Tris-glycine-SDS

Buffer used in SDS PAGE

Mercaptoethanol

Reagent used in sample buffer; Used to destroy/cleave disulfide bridges

Molecular weight

Data obtained from Native PAGE is the approximate _________ of the protein

Number of subunits and molecular weight

Data obtained from SDS PAGE is the approximate __________ and _____________

EDTA

Preservative that chelates divalent cations - reduces the activity of proteolytic enzymes

Tris-Cl

Acts as buffer

Glycerol

This makes the sample more dense than the sample buffer so the sample will remain at the bottom

Bromophenol blue

Dye that serves as a tracking dye for the progress of electrophoresis

SDS, TDT, and heat

These three are responsible for the actual denaturation of the sample

Negative charge

What gets added when the SDS breaks up the two- and three- dimensional structure of the proteins

Precipitation and aggregation

What will be the result of overloading that produces streaks and smears in electrophoresis

Isoelectric focusing (IEF)

Electrophoresis using IpH or pl as the basis; each protein will move until it reaches a position at the gel which the pH is equal to the IpH of the protein