Protein Electrophoresis
Protein Electrophoresis
Protein Electrophoresis
Prepared by
Eman Alshehri
Objectives
Separation of plasma protein and determine its
MWt.
components Amount
1.5 M Tris-HCL PH 8.8 2 ml
H2O 2.8 ml
components Amount
1.5 M Tris-HCL PH 6.8 1 ml
H2O 3 ml
10% SDS 80 µl
Acrylamide stock 1 ml
Running buffer pH 8.4
components Amount
Tris 15 g
Glycine 72 g
SDS 5g
Glycerol 1 ml
B- mercaptoethanol 0.5 ml
reparation
sults, all samples should be in identical, low ionic strength buffers.
of each sample with 10 μl of disruption buffer.
boiling water bath in for 2 min.
ases,brief boiling 3 min improves denaturation, but it may also cause the protein to precipi
Gels
SDS PAGE
Due to high density of binding of
SDS to proteins, the ratio
size/charge is nearly the same
for many SDS denatured
proteins. Hence proteins are
separated only by length of their
polypeptide chains (but not by
differences in charge).
Great separation. Allows
estimation of the size of
polypeptide chains
PAGE