Cell Fractionation Flashcards
Chop up fresh liver in an …… ……. Solution
Ice cold, buffer
What must the ice cold buffer solution be to the cells in the liver?
Isotonic
Step 3
Filter the mixture to remove debris.
What is the name of the machine the filtered mixture should be spun quickly in?
Centrifuge
What is found at the bottom of the test tube once spun
The denser part of the mixture is found at the bottom, this is known as sediment.
What is the name of the part of the mixture that is at the top once spun?
The supernatant
what happens to the supernatant after the first spin?
the supernatant is poured into a new tube and is spun again faster
What sediment is produced after the first spin?
The nuclei
What sediment is produced after the second spin?
The mitochondria
Why does the solution have to be ice-cold?
To reduce any enzyme activity as they may break down organelles
Why does the solution have to be isotonic?
To prevent any organelles bursting/shrinking due to osmosis gain/loss.
Why does the solution have to be buffered?
To prevent PH fluctuation which may alter structure/functioning of enzymes and/or organelles.
