NBT is prepared synthetically. The most common application for NBT is the detection of alkaline
phosphatase on western blots. NBT has also been used as a redox indicator for other enzymatic reactions including dehydrogenases, threonine deaminase, glucose-6-phosphate dehydrogenase, phosphofructokinase on polyacrylamide gels, oxidases on polyacrylamide gels, and pentose shunt
dehydrogenses.

Redox and halfwave potentials have been determined for NBT.  NBT has also been used as a colorimetric indicator of bacterial infection in blood samples.