BCIP/NBT One Component Membrane Alkaline Phosphatase (AP) Substrate

Store at temperatures from 2-25ºC. Do Not Freeze

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Stable for a minimum of 3 years from the manufactured date when properly stored between 2-25ºC. The substrate should not be frozen and should be protected from direct light by storing in amber bottles. Only high quality amber glass and plastic products should be used for storing aliquots.

This BCIP/NBT 1 component buffered alkaline phosphatase substrate (5-bromo-4-chloro-3-indolyl-phosphate and nitroblue tetrazolium) is a precipitating substrate used with the enzyme alkaline phosphatase designed for various membrane immunoassays but not recommended for microwell or immunohistochemical applications. Initially, the unreacted substrate should be colorless to light yellow in appearance. When this substrate system is reacted with alkaline phosphatase, an insoluble, permanent dark purple reaction product is obtained.

Leinco Technologies’ BCIP/NBT 1 Component Membrane Substrate is a highly sensitive, high performance formulation that produces a high signal to noise ratio with low membrane background. The outstanding shelf life of at least thirty six months for the BCIP/NBT 1 Component Membrane Substrate makes this reagent ideal for long term use of the same manufacturing lot.

BCIP/NBT 1 Component Membrane Substrate is a ready to use solution that needs no preparation or dilution. This product should be at room temperature (25°C) before use. Pour the necessary amount of substrate onto the blocked alkaline phosphatase conjugate labeled membrane to completely cover the membrane. The substrate will react with sites on the membrane containing phosphatase, producing an insoluble purple reaction product. The reaction should be monitored and read before the background color becomes too intense which may result in diminished contrast between the positive and background staining. To stop the reaction, rinse membrane with reagent quality water. Some wash solutions containing detergents may reduce color intensities. Dilution of the substrate is not recommended. To reduce the intensity of a reaction, it is recommended that the antibodies or conjugates be diluted.