Thermo Scientific Pierce Alkaline Phosphatase is purified calf-intestinal alkaline phosphatase (AP or alk-phos) enzyme for use in activity assays and conjugation to antibodies for ELISA and immunohistochemistry applications.
This purified calf intestinal alkaline phosphatase (CIP) is supplied in Tris buffer and 50% glycerol for use in protein research methods. The main applications for alkaline phosphatase in molecular biology and protein research are to remove 5'-phosphate groups from DNA or as a reporter system for immunoassays such as ELISA. For these latter methods, the enzyme is usually conjugated to specific primary or secondary antibodies and its activity is detected with a color-forming (or light-generating) substrate.
- Purified form – ready to dilute and conjugate without prior dialysis
- Concentrated – approximately 20mg/mL (lot-specific value reported)
- High specific activity – typically greater than 1600 units/mg (lot-specific value reported)
- Tris buffered solution – supplied in 5mM Tris, 5mM magnesium chloride and 0.1mM zinc chloride,
pH ~7.0 in 50% glycerol
One unit equals the amount of protein required to hydrolyze one micromole of p-nitrophenyl phosphate (PNPP) per minute at 25°C in a glycine buffer, pH 9.6.
- Bulman, A.S. and Heyderman, E. (1981). Alkaline phosphatase for immunocytochemical labeling: problems with endogenous enzyme activity. J. Clin. Pathol. 34, 1349-1351.
- Cordell, J.L., et al. (1984). Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). J. Histochem. Cytochem. 32, 219-229.
- Yolken, R.H. (1982). Enzyme immunoassays for the detection of infectious agents in body fluids: current limitations and future prospects. Rev. Infect. Dis. 4, 35-68.
Rerview of Immunohistochemistry (IHC)
Review of Immunodetection for IHC
Substrates for ELISA Detection
Substrates for Western Blot Detection
PNPP (p-Nitrophenyl Phosphate, Disodium Salt) Substrates