SensoLyte® pNPP Alkaline Phosphatase Assays – Improved Sensitivity


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.

Product

Catalog #

SensoLyte® pNPP Alkaline Phosphatase Assay Kit *Colorimetric* NEW

72146

SensoLyte® pNPP Alkaline Phosphatase ELISA Assay Kit *Colorimetric* NEW

72147-M

SensoLyte® pNPP Alkaline Phosphatase ELISA Assay Kit *Colorimetric* NEW

72147-R

SensoLyte® pNPP Protein Phosphatase Assay Kit *Colorimetric*

71105

SensoLyte® pNPP Secreted Alkaline Phosphatase Reporter Gene Assay Kit *Colorimetric* NEW

72144

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

Product

Catalog #

SensoLyte® FDP Alkaline Phosphatase Assay Kit *Fluorimetric*

71109

SensoLyte® FDP Protein Phosphatase Assay Kit *Fluorimetric*

71100

SensoLyte® FDP Secreted Alkaline Phosphatase Reporter Gene Assay Kit *Fluorimetric*

71108

SensoLyte® Luminescent Alkaline Phosphatase Assay Kit *Luminometric* NEW

72122

SensoLyte® Luminescent Alkaline Phosphatase ELISA Assay Kit *Luminometric* NEW

72123

SensoLyte® Luminescent Secreted Alkaline Phosphatase Reporter Gene Assay Kit *Luminometric* NEW

72145

SensoLyte® MFP Acid Phosphatase Assay Kit *Fluorimetric*

71231

SensoLyte® MFP Protein Phosphatase Assay Kit *Fluorimetric*

71104

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).