THE FRACTIONATION

OF A CELL'S CONTENTS
BY DIFFERENTIAL
CENTRIGUGATION
Reasol, Allona
Pendon, Shaldan
Yecyec, Rhovil
Bio 17.1 YB

CENTRIFUGATION

a common separation technique that

accomplishes separation based on the
the density and size differences in a
mixture of components.

2 types of centrifugal techniques:

a. Differential Centrifugation
b. Density- Gradient Centrifugation

DIFFERENTIAL
CENTRIFUGATION
a technique that isolates the
organelle at different density speeds.
this technique depends on a
principle:
as long as they are denser than the
surrounding medium, particles of
different size and shape travel toward
the bottom of a centrifugal tube ate
different rates when placed in a
centrifugal tube

STEPS:
Cell must be broken by mechanical
homogenization

Cell homogenate is subjected to

low speed centrifugation to pellet
the large particles for a short
period of time while smaller
vesicles (microsomes) are in
supernatant

Microsomes can be removed from

the supernatant by centrifugation
at higher-speeds

Ultracentrifuge, which generate

speeds of 75,000 revolution per
minute, equivalent to 500,000
times that of gravity

Figure 18.23
The medium is
composed of a
continuos
sucrosedensity
gradient, and
different
organelles
sediment until
they reach a
place in the
tube equal to
their own
density.

DENSITY-GRADIENT
CENTRIFUGATION

ultracentrifugation of substances in concentr

ated solutions of sucrose; at equilibrium

the medium exhibits aconcentration (hence

density) gradient increasing in the direction
of centrifugal force and the substances of
interest collect in layers at
the levels of their densities.

Size and density of some typical

organelles*
---------------------------------Organelle
diameter Density
(m)
(g/cm3)
---------------------------------Nuclei
5-10
1.4
Mitochondria 1-2
1.1
Ribosomes
0.02
1.6
Lysosomes
1-2
1.1
----------------------------------

Possible Errors

Differential centrifugation cannot yield

a pure preparation of an organelle

Too little then the organelles will not

be separated

Too much and even the molecules

can be broken

Application

Centrifugation can be used to any

cellular activities

It could be helpful to anyone who wishes

to study a specific type of cell organelles

Other Information

There are other techniques that can

purify a particular organelles such as
gravity sedimentation, simple sieves,
Isopycnic Centrifugation, Rate-Zonal
Centrifugation and etc.

Importance

The importance of the organelle being

isolated by this procedure can be used
in cell-free system to study a wide
variety of cellular activities including the,
synthesis of membrane-bound proteins,
the formation of transport of vesicles,
DNA synthesis and development of ionic
gradients.