Biomedical Engineering Reference
In-Depth Information
Fig. 4.2.
Pattern of immunoelectrophoresis.
A
Two antigen (
Ag
) samples,
one antiserum (
Ab
).
B
One antigen, two antisera. + Anode
4.11 Counterelectrophoresis
=
0, 1
8
A
barbital-acetate buffer, pH 8.2, I
Solutions/Reagents
B
0.5% agarose (w/v) in ddH
2
O
C
1.0% agarose with high endoosmosis (high EEO) (w/v) in
1:1 diluted Soln. A. Melt before use in a microwave oven or
a boiling water bath
A synonym for “counterelectrophoresis” is “crossing over elec-
trophoresis.”
Pre-coat slides with Soln. B (see Protocol 4.8.2) and prepare the
gel by pouring 0.2 ml
/
cm
2
molten Soln. C onto the slide placed on
a leveled surface. Be sure to cover the whole slide with a uniform
layer of agarose.
Punch at least two wells of 2- to 3-mm diameter using a Pasteur
pipet or a hypodermic needle. The wells are located in the middle
of the slide and should have a distance of 5 mm (Fig. 4.3).
Place the slide into a horizontal electrophoresis apparatus, fill
the tanks with Soln. A, and connect the small ends of the slide
to the electrode tanks by moistened filter paper wicks. Fill the
well directed towards the anode with antiserum dilution and pipet
antigen solution containing traces of bromophenol blue in that well
which is nearby the cathode.
2
·
n
=
1
(c
i
·
8
=
1
z
i
); c
i
, concentration of ion i, z
i
,chargeof
Ionic strength I
ion i
