Analysis of cell component - Cell Fractionation

Question 1

what is cell fractionation?

Answer 1

separates organelles and break open cells

Question 2

what are the three steps?

Answer 2

homogenisation
filtration
ultracentrifugation

Question 3

homogenisation - steps/conditions

Answer 3

the cells are broken up in a homogeniser/breaks down the cell membrane - leaves the organelles intact

conditions:

• ice cold
• isotonic
• buffer

Question 4

filtration - steps

Answer 4

solution is filtered by a gauze = separates cell/tissue debris

Question 5

Ultracentrifugation - steps

Answer 5

Cell fragments are poured into a tube in centrifuge - spun at low speed/ heaviest organelles form a pellet and light organelles stays suspended in the sol to form a supernatant/ supernatant is drained off and placed in the centrifuge at a higher speed/ process is repeated - pellet is replaced each time

Question 6

how are organelles separated ?

Answer 6

heaviest to lightest - in order of mass

nucleus - mitochrondrion - lysosome - endo r - ribosomes