Cell Fractionation Flashcards

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1
Q

What does cell fractionation mean?

A

Separating different parts and organelles of a cell, so they can be studied in detail.
Processes of cell metabolism (photosynthesis and respiration) have been studied this way/

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2
Q

What is the most common method of fractionating cells?

A

To use differential centrifugation.

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3
Q
  1. What is the first step in cell fractionation?
A

Cut the tissue in ice-cold isotonic buffer.

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4
Q

Why is the cold temperature needed to cut the tissue?

A

To slow enzyme reactions.

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5
Q

Why is the isotonic buffer needed to cut the tissue?

A

To stop osmosis, so organelles don’t burst.

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6
Q

Why is the buffer used to cut the tissue?

A

To stop pH change.

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7
Q
  1. What is the second step in cell fractionation?
A

Grind the tissue in a blender to break open cells.

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8
Q
  1. What is the third step in cell fractionation?
A

Filter.

Removing insoluble tissue like fat and plant cell walls.

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9
Q
  1. What is the filtrate now called?
A

A cell-free extract and its capable of carrying out most of the normal cell reactions.

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10
Q
  1. What is the fourth step in cell fractionation?
A

Centrifuge filtrate at low speed.
1000 x g for 10 mins.
This pellets nuclei which can be re-suspended

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11
Q
  1. What is the fifth step in cell fractionation?
A

Centrifuge supernatant at medium speed.
10,000 x g for 30 mins.
This pellets mitochondria and chloroplasts which can be re-suspended.

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12
Q
  1. What is the sixth step in cell fractionation?
A

Centrifuge supernatant high speed.
100,000 x g for 1 hour.
This pellets ER, golgi and other membrane fragments, which can be re-suspended.

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13
Q
  1. What is the seventh step in cell fractionation?
A

Centrifuge supernatant at VERY high speed.
300,000 x g for 3 hours.
This pellets ribosomes which can be re-suspended.

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14
Q
  1. What is the eighth step in cell fractionation?
A

Supernatant is now organelle- free cytoplasm.

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