Protein Spectometory

Question 1

What does the surface change of proteins depend on

Answer 1

PH

Question 2

What are reasons to purify proteins

Answer 2

-drug design
-disease mechanism
-commercial/ industrial production

Question 3

What purifying methods should be used based on a proteins size/shape

Answer 3

-centrifugation
-gel electrophoresis
-size-exclusion chromatography

Question 4

What purifying method/s should be used based on a proteins charge

Answer 4

-isoelectric focusing

Question 5

What purifying method/s should be used based on a proteins solubility

Answer 5

-salt fractionation

Question 6

What purifying method/s should be used based on a proteins ability to bind to molecules (ligands)

Answer 6

-Affinity chromatography

Question 7

What is salt fractionation

Answer 7

Separates proteins based on solubility at diff salt concs. Proteins w/ low solubility precipitate out of solution and other proteins will remain

Question 8

What is heat denaturation (fractionation)

Answer 8

Observes thermal stability of protein

Question 9

What is column chromatography

Answer 9

Separation technique of mixture passed through stationary phase material - different interactions with stationary phase and will be separated as they elute from the column

Question 10

What is gel electrophoresis

Answer 10

Separates molecules based on size and charge. Smaller proteins travel faster and a greater distance down the gel than larger proteins

Question 11

What are the three types of chromatographies (csbl)

Answer 11

-ion exchange chromatography - for charge
-size exclusion chromatography - size
-affinity chromatography - binding of ligands

Question 12

What is size exclusion chromatography (or gel filtration)

Answer 12

Separates molecules based on size. Smaller molecules get trapped in pores => elute slower
Large molecules pass through faster

Question 13

What is ion exchange chromatography

Answer 13

Stationary phase has beads with charged functional groups therefor when mixture passed through column, molecules with opposite charge are attracted to the bead and molecules with similar charge will pass through

Question 14

What is affinity chromatography

Answer 14

Sample passed through column and target molecule binds to ligand or tag while other molecules pass through. The target molecule then elute by changing conditions to disrupt interaction