Alkaline Phosphatase Substrate-Chromogen (BCIP/NBT)
1. Once tissue sections have been incubated with alkaline phosphatase, wash them with buffer thoroughly. 2. Wipe the glass to remove excess of buffer and add enough drops of the BCIP/NBT solution to cover the tissue sections. 3. Incubate for 10-30 minutes at room temperature. For the best results, look under the microscope for the signal development. Once desired signal to noise ratio is achieved, stop the reaction by washing the slides in the wash buffer.
Keep as concentrated solution, aliquot and store at 4ºC. Do not freeze.
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
Alkaline phosphatase, an enzyme derived from bovine intestinal mucosa, is often used as a label for in situ hybridization, immunohistochemistry, Southern and Northern hybridization and DNA sequencing. Detection of this enzyme requires the use of an insoluble colored substrate. GTX73326 is a new stable, BCIP/NBT solution in ready to use form. It produces a blue colored end product at the positive sites which gives a good contrast with red nuclear fast counter stain. Specimens stained in BCIP/NBT can not be dehydrated in ethanol and hence should be mounted in aqueous based mounting medium.