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BCIP/NBT MEMBRANE
SUBSTRATES
BCIP (5-Bromo-4-Chloro-3'-Indolyphosphate p-Toluidine
Salt) and NBT (Nitro-Blue Tetrazolium Chloride)
are chemical compounds used in immunoblotting
for the sensitive detection of alkaline
phosphatase (AP). NBT serves as the oxidant and
BCIP is the alkaline phosphatase substrate. BCIP/NBT
naturally forms a bluish-purple precipitate over
time; however, alkaline phosphatase speeds up
the process 1000-fold. BCIP binds very tightly
in the alkaline phosphatase active site, but
when NBT reacts with BCIP, it is released from
the enzyme and the colored precipitate forms.
BioFX has developed two unique one component
ready to use BCIP/NBT substrates for the
detection of alkaline phosphatase in membrane
applications. These two formulations, available
in blue or purple, provide crisp detail and low
background.
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