Electrophoresis is a separation technique that is based on the movement of charged particles in an electric field
The term electrophoresis was coined from a greek work “Phoresis” which means “Being carried away”
Hence literal meaning of the word electrophoresis means “to carry with electricity.”
an analytical method used to separate components of a protein mixture based on size.
Principle : A charged molecule will migrate in an electric field towards an electrode with opposite sign.
Polyacrylamide gels are characterized by two parameters: total monomer concentration (%T, in g/100 ml) and weight percentage of cross linker (%C).
%T -the relative pore size of the resulting polyacrylamide gel
higher %T - smaller pores.
The practical ranges for monomer concentration are stock solutions of 30-40%
2. Electrophoresis
Electrophoresis is a separation technique that
is based on the movement of charged particles
in an electric field
The term electrophoresis was coined from a
greek work “Phoresis” which means “Being
carried away”
Hence literal meaning of the word
electrophoresis means “to carry with
electricity.”
3. PAGE Introduction
an analytical method used to separate
components of a protein mixture based on
size.
Principle : A charged molecule will migrate in
an electric field towards an electrode with
opposite sign.
4.
5. Native PAGE
Separation is based upon charge,size
and shape of macromolecules.
Useful for separation and purification
of mixture of proteins
This was the original mode of
electrophorosis
6. POLYMERISATION
Initiated by free radical catalysis
Polymerisation of acrylamide monomer and
bisacrylamide comonomer
Head to tail fashion
Fairly well defined structure
7.
8. Polyacrylamide gels are characterized by two
parameters: total monomer concentration (%T,
in g/100 ml) and weight percentage of cross
linker (%C).
%T -the relative pore size of the resulting
polyacrylamide gel
higher %T - smaller pores.
The practical ranges for monomer
concentration are stock solutions of 30-40%
9. Advantages
Estimation of molecular weight of proteins and
nucleic acids
Determination of subunit structure of proteins
Purification of isolated proteins
Monitoring changes of protein content in
body fluids
