LOW AND HIGH VOLTAGE
ELECTROPHORESIS
Movement of charged particles under the influence of the electric
current
CONTENT
INTRODUCTION
LOW VOLTAGE ELECTROPHORESIS
• INSTRUMENTAION
• APPLICATION
HIGH VOLTAGE ELECTROPHORESIS
• INSTRUMENTATION
• APPLICATION
INTRODUCTION
• Separations technique that is based on the
mobility of ions in an electric field.
• Types of electrophoresis:
GEL LOW AND HIGH
ELECTROPHORESIS VOLTAGE
ELECTROPHORESIS
CAPILLARY CONTINOUS
ELECTROPHORESIS ELECTROPHORESIS
LOW VOLTAGE
ELECTROPHORESIS
INSTRUMENTATION [GENERAL
APPARATUS]
• It consists basically of two items a power pack
and an electrophoresis unit.
POWER PACK
ELECTROPHORESIS UNIT
Electrodes
Buffer reservoir
Support for electrolysis medium
A transparent insulating cover.
LOW VOLTAGE
ELECTROPHORESIS
The apparatus commonly used for low voltage
paper electrophoresis is of two types:
HORIZONTAL STRIP
VERTICAL STRIP
HORIZONTAL STRIP
Principle: To separate any ionic substances
VERTICAL STRIP
APPLICATION OF LOW VOLTAGE
ELECTROPHORESIS
• In examination of biological and clinical
specimens for amino acids and proteins.
• Use for separation of proteins and other high
molecular weight substance.
HIGH VOLTAGE
ELECTOPHORESIS
INSTRUMENTATION(GENERAL
APPARATUS)
• The equipment consists basically of two items a
power pack and an electrophoresis unit.
POWER PACK
ELECTROPHORESIS UNIT
Electrodes
Buffer reservoir
Support for electrolysis medium
A transparent insulating cover.
Cooling Plate.
HIGH VOLTAGE ELECTROPHORESIS
• The apparatus commonly used for high voltage
paper electrophoresis is of two types:
ENCLOSED STRIP
IMMERSED STRIP
ENCLOSED STRIP
• In principle, paper electrophoresis is simpler
than paper chromatography.
• There are two features of an ion which are
most pertinent to its electrophoretic
behavior; the size of the ion and the size of
the charge it carries.
• These features oppose each other; the bigger
the size the slower the ion will travel; the
bigger the charge the faster it will go.
IMMERSED STRIP METHOD
WORKING:
• In this method the sample is applied to a strip
of filter paper maintained with buffer solution
and the strip is immersed in a large volume of
buffer- immiscible, electrically insulating and
volatile cooling liquid.
• Apparatus can also be designed to accumulate
cooling liquids which have a higher density than
that of buffer solution.
APPLICATION OF HIGH VOLTAGE
ELECTROPHORESIS
• High voltage electrophoresis has been
successfully used in the separation of low
molecular weight substance such as amino acids,
peptide, Indoles, sugars, Purines, Pyrimidines,
amines and inorganic ions.
• The amino acid composition of two unknowns can
be determined.
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