pNPP Phosphatase Assay Kit
Datasheet PDF  MSDS/Bulk Order


Application
For quantitative determination of phosphatase activity.

Key Features
High sensitivity and wide linear range. The detection limit is generally 3 ng phosphatase or below.

Homogeneous and simple procedure. No wash or reagent transfer steps are involved. The assay can be completed within 30 minutes.

Robust and amenable to HTS. All reagents are compatible with high-throughput liquid handling instruments.

Method
OD405nm

Samples
Protein phosphatases

Species
All

Procedure
30 min

Size
500 tests

Detection Limit
3 ng

Shelf Life
12 months

More Details
p-nitrophenyl phosphate (pNPP) is a chromogenic substrate for most phosphatases such as alkaline phosphatases, acid phosphatases, protein tyrosine phosphatases and serine/threonine phosphatases. The reaction yields para-nitrophenol, which becomes an intense yellow soluble product under alkaline conditions and can be conveniently measured at 405 nm on a spectrophotometer. This homogeneous "mix-and-measure" assay involves simply adding a single reagent to the phosphatase and measuring the product formation on an absorbance reader. The assay can be conveniently performed in cuvettes, tubes or multi-well plates at either room temperature or 37°C.

1. I would like to run a positive control, to compare to her data. Will the Sigma enzyme (catalog # P6772, bovine alkaline phosphatase) work with your assay kit as a positive control? (i.e. pNPP substrate + assay buffer = units of activity as compared to the known units of activity in the Sigma enzyme)



The Sigma enzyme should work with our assay kit. As indicated in our assay protocol, the customer needs to prepare their own enzyme buffer (100 mM Tris-HCl (pH 8.6), 10 mM MgCl2 or similar buffers) to raise the pH to 8.6 for an alkaline phosphatase.

2. Does your pNPP kit measure the units of activity or does it measure the moles of pNPP liberated/ minute?



Out kit measures the moles of pNPP liberated per minute as a unit for enzyme activity.

For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions
1. Huxtable AG et al (2009). Tripartite purinergic modulation of central respiratory networks during perinatal development: the influence of ATP, ectonucleotidases, and ATP metabolites. J Neurosci. 29(47):14713-25. Assay: Phosphatase in Human cells (Pubmed).

2. Lee SW et al (2008). The Xanthomonas oryzae pv. oryzae PhoPQ two-component system is required for AvrXA21 activity, hrpG expression, and virulence. J Bacteriol. 190(6):2183-97. Assay: Phosphatase in Human mesenchymal stem cells (Pubmed).

3. Oborna I et al (2010). Increased lipid peroxidation and abnormal fatty acid profiles in seminal and blood plasma of normozoospermic males from infertile couples. Hum Reprod. 25(2):308-16. Assay: Phosphatase in Fish marine teleost follicles, eggs (Pubmed).

4. López-Vales R et al (2010). Fenretinide promotes functional recovery and tissue protection after spinal cord contusion injury in mice. J Neurosci. 30(9):3220-6. Assay: Phosphatase in Mouse cell supernatant (Pubmed).

5. Monick, MM et al (2006). Active ERK Contributes to Protein Translation by Preventing JNK-Dependent Inhibition of Protein Phosphatase. J. Immunol. 177: 1636–1645. . Assay: Phosphatase in Bacteria cells (Pubmed).

6. Sawaguchi S et al (2006). Identification of two forms of vitellogenin-derived phosvitin and elucidation of their fate and roles during oocyte maturation in the barfin flounder, Verasper moseri. Zoolog Sci. 23(11):1021-9. Assay: Phosphatase in Zebrafish serum (Pubmed).

7. Nakano Y (2007). Novel function of DUSP14/MKP6 (dual specific phosphatase 14) as a nonspecific regulatory molecule for delayed-type hypersensitivity. British J. Dermatology 156 (5): 848–860. Assay: Phosphatase in Chicken intestinal tissue (Pubmed).

To find more recent publications, please click here. To find more recent publications, please click here.
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pNPP Phosphatase Assay Kit
Catalog No: POPN-500
Price: $159    Qty:
For orders of 10 or more kits, please contact us at 1-510-782-9988 x 1 for best pricing.

Shipping: RT
Shipment: Same Day for order by 2pm PST
Delivery: 1-2 days (US), 3-6 days (Intl);
Storage: 4, -20°C

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