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TapeshwarYadav
(Lecturer)
BMLT, DNHE,
M.Sc. Medical Biochemistry
The technique of paper electrophoresis is simple and
inexpensive and requires only micro quantities of plasma
for separation.
The support medium is a filter paper
The electrophoresis apparatus in its simplest form
consists of two troughs to contain buffer solution,
through which electric current is passed.
Frequently used in isolating proteins, amino acids and
oligopeptides.
IntroductionIntroduction
ProcedureProcedure
1) A long strip of filter paper is moistened with a suitable
buffer solution of the desired p H and the sample is applied
transversely across the central part of the strip.
2) Ends are fixed to dip in buffer solutions in two troughs
fitted with electrodes.
3)Electric field of about 20 volts/cm is established.
4)The charged particles of sample migrate along the strip
towards respective electrodes of opposite polarity,
according to net charges, sizes and interactions with the
solid matrix.
Contd…Contd…
5)Homogeneous group of particles migrate as a separate
band
6)The electrophoresis is carried out for 16-18 hours.
7) Separated Proteins are fixed to a solid support using a
fixative such as Acetone or Methanol
8)Proteins are stained (bromophenol blue) to make
them visible
9) The separated proteins appear as distinct bands
10)Drawback-long time interval and blurring of margins
The different fractions appear as blue coloured
bands across the filter paper starting from the
moving boundary backwards.
 If a quantitative estimation is required for each
fraction, the bands may be carefully cut and eluted,
or the bands may be scanned optically in a
densitometer.
 In human plasma five different bands can be
identified on paper electrophoresis
ObservationObservation
Paper Electrophoresis Guide
Paper Electrophoresis Guide
Paper Electrophoresis Guide

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Paper Electrophoresis Guide

  • 2. The technique of paper electrophoresis is simple and inexpensive and requires only micro quantities of plasma for separation. The support medium is a filter paper The electrophoresis apparatus in its simplest form consists of two troughs to contain buffer solution, through which electric current is passed. Frequently used in isolating proteins, amino acids and oligopeptides. IntroductionIntroduction
  • 3.
  • 4.
  • 5. ProcedureProcedure 1) A long strip of filter paper is moistened with a suitable buffer solution of the desired p H and the sample is applied transversely across the central part of the strip. 2) Ends are fixed to dip in buffer solutions in two troughs fitted with electrodes. 3)Electric field of about 20 volts/cm is established. 4)The charged particles of sample migrate along the strip towards respective electrodes of opposite polarity, according to net charges, sizes and interactions with the solid matrix.
  • 6. Contd…Contd… 5)Homogeneous group of particles migrate as a separate band 6)The electrophoresis is carried out for 16-18 hours. 7) Separated Proteins are fixed to a solid support using a fixative such as Acetone or Methanol 8)Proteins are stained (bromophenol blue) to make them visible 9) The separated proteins appear as distinct bands 10)Drawback-long time interval and blurring of margins
  • 7. The different fractions appear as blue coloured bands across the filter paper starting from the moving boundary backwards.  If a quantitative estimation is required for each fraction, the bands may be carefully cut and eluted, or the bands may be scanned optically in a densitometer.  In human plasma five different bands can be identified on paper electrophoresis ObservationObservation