Alkaline Phosphatase Enhancer (ab671)

To use as signal enhancer in conjunction with alkaline phosphatase based immunostaining systems.

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 generation of an insoluble colored reaction end product. The stronger the signal, the better the staining. This is a new stable solution to increase the signal generated by alkaline phosphatase several fold. A single treatment of tissue sections with enhancer before the treatment with substrate/chromogen enables the visualization of difficult to localize antigens. This enhancer works well with Fast Red but can be used with BCIP/NBT and BCIP/INT also. Specimens stained with the above substrates/chromogens can not be dehydrated in ethanol and hence should be mounted in aqueous based mounting medium.

Protocol:

i) Once tissue sections have been incubated with streptavidin-alkaline phosphatase, wash them with buffer thoroughly.

ii) Wipe the glass to remove excess of buffer and add enough drops of the alkaline phosphatase enhancer solution to cover the tissue sections. Alternately, pour alkaline phosphatase enhancer solution in a coplin jar and dip the slides for 1 minute.

iii) Incubate for 1 minute at room temperature.

iv) Drain excess alkaline phosphatase enhancer and add substrate/chromogen onto the tissue sections without any wash between the two steps.

Precautions:Avoid contact with clothes and exposed skin. If accidentally contacted, flush with tap water immediately. Do not take internally.