Molecular Separations Flashcards
Purifying, separating and identifying mixtures containing proteins.
Proteins can be separated by their differing properties of (3)…
Size, charge and affinity
Chromatography consists of a mobile phase and a…
Stationary phase
The two main types of chromatography are…
- Thin-layer (solubility)
* Column chromatography (affinity for stationary phase)
When the molecules leave the chromatography column, this is known as…
Eluting
The sample is known as…
Eluate
Gel electrophoresis is used to…
Analyse proteins or DNA/RNA 🔍
“PAGE” stands for…
PolyAcrylamide Gel Electrophoresis
Native PAGE allows the separation of proteins by the combination of…
Size, charge and shape
The downside to native PAGE is that it…
Cannot distinguish between the effects of size, charge or shape
SDS-PAGE enables proteins to be separated by…
Size only
In SDS-PAGE, eluate added to the wells moves through…
The stacking gel, then the running gel
SDS denatures proteins and adds negative charge…
This makes the results shape and charge independent (dependent only on size)
Heat the protein to 95°c with SDS and ? to disrupt SS bonds and denature
Mercaptoethanol (reduces disulfide bonds)
In SDS-PAGE the migration of a protein is proportional to…
the log of the molecular weight
IEF stands for…
Iso-electric focussing
