The invention discloses an
isoelectric focusing electrophoretic method of hemopoietin. A BIO-RAD 111 type
electrophoresis apparatus is used to carry out
isoelectric focusing electrophoresis. Each 5 mLof glue comprises 1 mL of
acrylamide monomer solution (25 wt%), 3.33 mL of
urea (10.5 M), 0.412 mL of
cane sugar (60wt%), 0.183 mL of amphoteric
electrolyte solution (40 wt%, pH 2-4), 0.075 mL of amphoteric
electrolyte solution (40 wt%, pH 3-10), and a catalyst solution, wherein the catalyst solution is mixed in advance and comprises 25 [mu]L of FMN-Na (0.1 wt%), 7.5 [mu]L of AP (10 wt%), and 1.5[mL] of TEMED. Before sample injection, pre-
electrophoresis is performed for 10 minutes under a
voltage of 200 V.
Electrophoresis is performed through following phases: first phase, 50 V, 30 minutes;second phase, 100 V, 60 minutes;
third phase, 200 V, 30 minutes; fourth phase, 450 V, 60 minutes; and fifth phase, 590 V, until the current is zero. The method can separate the hemopoietin charge isomers with different isoelectric points, and has the advantages that the requirement on the upper limit of
voltage is not high and no cooling apparatus is needed.