Bio-Rad Immun-Blot® PVDF Membrane - Manual (Page 4)

can be added to the transfer buffer without
affecting the binding of the protein to PVDF.
The results are consistently clean, easy to read
blots. The physical strength of Immun-Blot
PVDF membrane means that it will not crack or
tear in common handling, and holds up under
repeated strip and reprobing applications.
Section 2
Membrane Wetting
Immun-Blot PVDF membrane can be used
as a direct replacement for the membrane cur-
rently being used in your blotting protocol. No
changes are required in the procedure, but the
special steps given below are required to pre-
pare the membrane for blotting. The hydropho-
bicity of PVDF makes it impossible to wet the
membrane with aqueous solutions. Methanol or
an alternative organic solvent is required to pre-
wet the membrane prior to equilibration in
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much protein as possible, a western blot requires
good signal retention with very low background.
In order to provide the best possible membrane
for each technique, Bio-Rad offers two grades
of PVDF, Immun-Blot PVDF for western blot-
ting, and Sequi-Blot PVDF for protein sequenc-
ing.
Immun-Blot PVDF
The purpose of a blotting membrane is to
deliver good signal results while resisting back-
ground and non-specific binding. Immun-Blot
PVDF Membrane is ideal for chemiluminescent
and colorimetric western blots because it retains
target protein very strongly, 140–150 µg pro-
tein/cm
2 membrane, but resists background that
can obscure high sensitivity detection. Immun-
Blot PVDF membrane retains proteins in any
transfer format; tank blotting, semi-dry blotting,
and dot blotting all deliver excellent results. For
proteins that are difficult to transfer, 0.1% SDS
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