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Introduction – Hoefer PR625 Immunoblot User Manual

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Introduction

Transfer of proteins or nucleic acids, fraction-
ated by gel electrophoresis, to an immobilizing
membrane surface increases the sensitivity of
a wide range of detection methods, reduces
time of analysis and makes sequential probing
possible. In particular it has made possible use
of immunological procedures that are not practi-
cal to carry out in a gel. To assay material on a
sheet of membrane with different probes simul-
taneously commonly requires first cutting the
membrane into a number of parallel strips. This
procedure however destroys the correspondence
between different lanes or positions on a gel.

The ImmunoBlot

®

and ImmunoBlot XL

preserve this correspondence while minimiz-
ing the volume of detection reagents needed for
individual lanes. Two clear plastic plates, one
smooth and one with channels, clamp a 15 ×
15 cm transfer membrane to define and separate
the original gel lanes into individual channels.
These separate channels allow for the use of
different detection reagents (primary antibodies,
conjugated secondary antibodies, antigens and/
or reaction substrates) in each channel.

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