AnaSpec’s widely used and cited (29 literature citations) SensoLyte® pNPP Alkaline Phosphatase Assay kits now contain a specially formulated pNPP substrate. This new substrate outshines the previous substrate in that it is:
• Ready to use – no need to reconstitute pNPP powder
• Improved sensitivity – detects as low as 0.02 ng alkaline phosphatase
• Improved stability – stores neat at 4C for one year

Changes in alkaline phosphatase level and activity are involved in a variety of physiological and pathological events, such as bone development,¹ bone-related diseases,² gestation related diseases,³ inflammatory bowel disease,⁴ post-parathyroidectomy stage,⁵ and drug toxicity.⁶ Alkaline phosphatase is widely used in ELISA for conjugation with secondary antibody and as a reporter for gene expression studies.

These SensoLyte® pNPP Alkaline Phosphatase Assay kits provide a convenient colorimetric assay for detecting alkaline phosphatase in biological samples and in ELISA with alkaline phosphatase conjugated secondary antibody or streptavidin by using colorimetric pNPP (p-Nitrophenyl phosphate) phosphatase substrate. Upon dephosphorylation, pNPP turns yellow and can be detected at absorbance=405 nm.

The SensoLyte® pNPP Alkaline Phosphatase ELISA Assay kits provide an AP-conjugated secondary antibody – goat anti-mouse (cat# 72147-M) or goat anti-rabbit (cat# 72147-R). AnaSpec also offers a pNPP assay kit for secreted alkaline phosphatase reporter assay, the SensoLyte® pNPP Secreted Alkaline Phosphatase (SEAP) Reporter Gene Assay Kit. A luminometric SEAP kit is also available.

Figure 1. Detection of alkaline phosphatase with the SensoLyte® pNPP Alkaline Phosphatase Assay Kit (Cat# 72146)

Alkaline phosphatase enzyme at each dilution was mixed with pNPP substrate, and then incubated at room temperature for 1 hr. Absorbance was recorded at 405 nm (Ultra Microplate Reader EL808, Bio-Tek Instruments, Inc). This assay is able to detect as low as 0.02 ng of alkaline phosphatase.

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Table 1. New pNPP formulation alkaline phosphatase assay kits.

Table 2. Fluorimetric and luminometric SensoLyte® Alkaline and related phosphatase assay kits.

References

1. Kotobuki, N. et al. Cell Transplant. 13, 377 (2004).
2. Wyckoff, MH. et al. J. Clin. Endocrinol. Metab. 90, 1233 (2004).
3. Boronkai, A. et al. J. Clin. Pathol. 58, 72 (2005).
4. Sanchez, M. et al. Biochem. Pharmacol. 68, 2317 (2004).
5. Morrone, LF. et al. Ann. Ital. Med. Int. 19, 189 (2004).
6. Papaldo, P. et al. Cancer Invest. 22, 650 (2004).