lab procedures 9/17

Question 1

what is the first step for handling proteins

Answer 1

purification

Question 2

what does purification do

Answer 2

get rid of proteins you don’t want

Question 3

what is an assay

Answer 3

locate and quantify the protein of interest in the extract and in each of the fractions

Question 4

what is crude extract

Answer 4

biological source of the desired protein is homogenized to break the cells into parts that can be separated

Question 5

what is subcellular fractionation and what are the types

Answer 5

separating the crude extract into fractions containing specific organelles, or soluble proteins

• rate zonal centrifugation
• equilibrium density centrifugation

Question 6

what is rate zonal centrifugation

Answer 6

• separates based on sedimentation coefficient
• based on size and density
• first step in subcellular fractionation

Question 7

equilibrium density centrifugation

Answer 7

organelle fraction separates based on density and stops at density it is

Question 8

what is enzyme activity formula

Answer 8

units = micromoles product / minute

Question 9

what is specific activity (purity) formula

Answer 9

units/mg of protein

Question 10

what does increased specific activity indicate

Answer 10

more pure enzyme