3.1. Protein Analysis Technique: Seperation & Purification

Question 1

SDS

Answer 1

Sodium dodecyl sulfate

1. Amphipathic detergent with an anionic tail and a lipophilic tail
2. Used to denature and dissociate from each other
3. Confers a negative charge on the protein in turn masking the proteins intrinsic charge
4. SDS treated proteins have similar mass to charge ratios and shapes
5. During PAGE the rate of protein migration is determined by molecular weight where all proteins migrate towards the anode

Question 2

PAGE

Answer 2

poly-acrylamide gel electrophoresis

1. Support medium

Question 3

Rf

Answer 3

Relative migration distance of protein

Question 4

IEF

Answer 4

Isoelectric focusing

Question 5

Isoelectric point

Answer 5

The point where the overall charge on the protein is zero (a neutral charge)

Question 6

Ampholytes

Answer 6

Complex mixtures of synthetic poly amino-polycarboxylic acids

Question 7

SDS-PAGE

Answer 7

Proteins focused in the 1st dimension are resolved in a perpendicular plane by conventional SDS-PAGE

Question 8

Chromatography

Answer 8

Fundamental component of most (if not all) recombinant protein purification