H922 34: Cell Biology Outcome 4: Investigating the biochemical properties of the plasma

membrane

INVESTIGATING THE BIOCHEMICAL PROPERTIES OF THE PLASMA

MEMBRANE
AIM
Investigate the composition of the plasma membrane by examining the effect
of temperature and detergent on cell membrane structures

INTRODUCTION
The cell membrane is composed of a phospholipid bilayer with proteins
dispersed throughout. One of the main functions of the cell membrane is to
regulate what can enter and leave the cell. When the cell membrane is
damaged the contents of the cell can leak out into the surrounding fluid.
In this experiment we will be using discs of red cabbage to examine the
properties of the cell membrane. Inside red cabbage cells there are vacuoles
which contain a red pigment which belong to a class of molecules called
anthocyanins. When the cell membrane is intact the pigment remains inside
the cell. However when the cell membrane is damaged the pigment leaks out
of the cell and into the surrounding environment which will then take on a red
colour.
The greater the damage to the membrane the greater the amount of dye that
leaks out. This can be monitored using a colorimeter which will measure the
intensity of the red dye allowing us to compare different treatments to see
their effect on the integrity of the plasma membrane.
In this investigation we will be examining the effect of temperature and
detergent on the plasma membrane. At high temperatures proteins in the
plasma membrane will become denatured. This will affect the integrity of the
plasma membrane and allow the cell contents to leak out. We will also
examine the effect of detergent on the cell membrane. Biological detergents
are commonly used to disrupt the bipolar lipid membrane of cells in order to
release and solubilise membrane-bound proteins. This treatment will disrupt
Adapted from Nuffield Foundation Protocol

H922 34: Cell Biology Outcome 4: Investigating the biochemical properties of the plasma
membrane

the cell membrane and allow the cell contents (including the red pigment) to
leak out of the cell.

MATERIALS AND METHODS

Safety Notes
Take care when using the cork borer. Only use it on the white tile provided
and not on the bench surface.
Take care when using the water baths. Your test tubes will be hot when you
take them from the water bath at the end of the experiment.
The dye released from the red cabbage will stain but it is not harmful.
Equipment List
Per group:

4 cm3 2% SDS solution

White Tile

Red cabbage leaf

Cork Borer No. 4

Beaker

Per Class

9 test tubes

Water Baths set to 30, 40, 50 and

Test tube rack

60 C.

Marker Pen

Colorimeter set to 550 nm

2 mL graduated pipette + filler

Cuvettes

Part 1 Preparing the Discs of Red Cabbage

1. Take a leaf of red cabbage and place on a white tile.
2. Cut at least 50 discs from the leaf using the cork borer and place in a
beaker of cold tap water.
Adapted from Nuffield Foundation Protocol

H922 34: Cell Biology Outcome 4: Investigating the biochemical properties of the plasma
membrane

3. Leave the discs to soak until you are ready to use them. This ensures
any dye that leaked out during the cutting process is removed.
4. You may need to replace the water until it remains clear.
Part 2 - Effect of Detergent
1. Label 5 test tubes 2%, 1%, 0.5%, 0.25% and 0%.
2. In all test tubes except the one labelled 2% place 2 cm 3 of distilled
water.
3. Add 2 cm3 of 2% SDS to the tube labelled 2% using a graduated 2 mL
pipette.
4. Add 2 cm3 of 2% SDS to the tube labelled 1% and mix gently. (The total
volume in this tube is now 4 cm3)
5. Take 2 cm3 of the solution from the 1% tube and add it to the 0.5% tube
and mix gently.
6. Take 2 cm3 of the solution from the 0.5% tube and add it to the 0.25%
tube and mix gently.
7. Take 2 cm3 of the solution from the 0.25% tube and discard it. You
should now have 2 cm3 of solution in each test tube.
8. Add 5 red cabbage discs to each tube.
9. Incubate the tubes for 20 minutes at room temperature.
10. After 20 minutes observe the tubes for any red dye that has leaked
from the discs and record your results in a table.
11. Use the colorimeter to measure the amount of dye that has leaked from
the cells and record your results in a table.
Part 3 - Effect of Temperature
1. Label 4 test tubes with your initials and 30, 40, 50 and 60.
2. Place 2 cm3 of water in each test tube using the 2 mL graduated
pipette.
3. Add 5 red cabbage discs to each test tube.
4. Place the test tube in the appropriate water bath for 20 minutes.
5. After 20 minutes observe the tubes for any red dye that has leaked
from the discs and record your results in a table.
Adapted from Nuffield Foundation Protocol

H922 34: Cell Biology Outcome 4: Investigating the biochemical properties of the plasma
membrane

6. Use the colorimeter to measure the amount of dye that has leaked from
the cells and record your results in a table.

TECHNICAL NOTES
2% SDS
2 g of SDS added to 100 cm3 of water. Stir to dissolve. Take care with SDS
powder. Do not breath the dust. Don not store in the cold.
If red cabbage is not available beetroot can be used instead. Beetroot must
be raw, not cooked. Use a size 4 cork borer and cut with care using a cutting
board. Cut enough cores to make eight 2 cm lengths per working group.
Leave the cores overnight in a beaker of distilled water. The pigment from any
cells that have been cut by the cork borer will leak into the water. Rinse away
any pigmented water in the morning and replace with fresh water.
Beetroot cells contain pigments called betalains that give the tissue its dark
purple-red colour. The pigment is contained in the cell vacuole.

Adapted from Nuffield Foundation Protocol