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Biochemical Methods I Notes

Pharmacology Notes > BIOL10212 Biochemistry Notes

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Biochemistry - Lecture 11 (05/03/2018)

Biochemical Methods I
Isolation of Proteins
Isolation from cells and tissues to molecules:

Organisms are an extremely mixture of chemicals and if Biochemists wish to identify which chemicals they are made of then methods of isolation are required.

Stages in protein purification 1) The physical disruption of cells or tissues.
2) Differential centrifugation to isolate cellular organelles.
3) Separation of proteins on the basis of: Solubility, Size, Charge, Hydrophobicity,
Affinity for ligands.

Separation of organelles by Differential Centrifugation

The acceleration (a) is often given as: g (the force of gravity).
g = 9.8 ms-2 = 9.8 N kg-1

a = - ω2r a = acceleration (ms-2)
ω = angular velocity (in radians/sec = s-1)
r = radius (m) ω=

2 π rpm 60

ω = angular velocity (in radians/sec = s-1)
rpm = revolutions per minute.

RCF =
RCF =

a g

=

rω2 g

𝒂

RCF = Relative Centrifugal Force.
ω = angular velocity (in radians/sec = s-1)
g = acceleration due to gravity (9.81ms-2).
a = acceleration (ms-2).
r = radius (m).

The Calculation of Acceleration

Usually, the average distance is used for the radius (r)
parameter in the calculation. For example, for this rotor, b/

2.

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