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Western blotting using the ProtoBlot® II AP System with Stabilized
Substrate
Gregg Larson
Promega Corporation
The ProtoBlot® II AP Systems with Stabilized Substrate are designed for
the rapid, sensitive immunodetection of proteins or other macromolecular antigens
immobilized on nitrocellulose or polyvinylidene fluoride (PVDF) membranes. The ProtoBlot
II AP Systems improve upon the simplicity of the original ProtoBlot Western AP Systems by
incorporating the premixed, ready-to-use Western BlueTM Stabilized Substrate
for Alkaline Phosphatase. When ProtoBlot II is used in Western blotting, less than 50pg of
antigen can be detected after 10 minutes of color development, while overnight development
increases the sensitivity further. Information is provided on the sensitivity levels of
the ProtoBlot II System under a variety of conditions.
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