Alkaline phosphatase (ALP) is a phosphomonoester hydrolase that is commonly used as a conjugating enzyme in biological research. A wide variety of substrates have been developed to assay its activity. In this study, we developed an ALP assay method utilizing merocyanine 700 (MC700) based substrate MC700 phosphate (MC700p). MC700 is a near-infrared fluorescent merocyanine dye, and has excitation/emission maxima at 686 nm/722 nm in ALP assay buffer. Upon hydrolysis by ALP, MC700p is converted to MC700. The fluorescence of MC700 is dependent on the pH and detergent concentration in the buffer. The fluorescence signal produced by MC700p hydrolysis is linearly related to the ALP amount and substrate concentration. A stop solution containing EDTA could be used to stop the ALP/MC700p reaction. It was also demonstrated that MC700p could substitute pNpp as the ALP substrate in a commercial 17β-Estradiol enzyme immunoassay kit.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.talanta.2011.02.035 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Unusual Association of Partial Fanconi Syndrome and Tumor-Induced Osteomalacia Revealed by Multiple Vertebral Fractures.
Calcif Tissue Int
January 2025
Division of Bone Diseases, Department of Medicine, Geneva University Hospitals, Geneva, Switzerland.
Tumor-induced osteomalacia (TIO) is a rare acquired paraneoplastic syndrome caused by a mesenchymal tumor secreting a phosphaturic hormone called FGF23. Patients present with bone pain, fragility fractures and muscle weakness. Biochemical results show hypophosphatemia, raised serum alkaline phosphatase and reduced calcitriol.
View Article and Find Full Text PDFSeminal plasma metabolomics and sperm lipidomics profiles of bull semen with different total progressive motile sperm count.
J Anim Sci
January 2025
Henan International Joint Laboratory of Nutrition Regulation and Ecological Raising of Domestic Animal, College of Animal Science and Technology, Henan Agricultural University, Zhengzhou, China.
Total progressive motile sperm count (TPMSC) is a reliable index of fecundity evaluation of bull semen. It is an important determinant frozen semen yield and conception rate of females artificially inseminated. Seminal plasma metabolites and sperm lipids are closely related to sperm survival and motility, but their relationship with TPMSC is not well known.
View Article and Find Full Text PDFOscillatory fluid flow enhanced mineralization of human dental pulp cells.
Front Bioeng Biotechnol
January 2025
Department of Preventive Dentistry, Division of Pediatric Dentistry, Faculty of Dentistry, Naresuan University, Phitsanulok, Thailand.
The purpose of this study is to evaluate the optimum frequency of oscillatory fluid flow (OFF) for increasing osteogenesis in human dental pulp cells (DPCs) in an incubating rocking shaker. DPCs from 3 donors were cultured in an osteogenic induction medium (OIM) and mechanical stimulation was applied using an incubating rocking shaker at frequencies of 0 (control), 10, 20, 30, and 40 round per minute (RPM) for 1 h/day, 5 days/week. Cell proliferation was measured using total protein quantification, and osteogenic activity was measured by alkaline phosphatase (ALP) activity, calcium deposition, and collagen production on days 7, 14, and 21 of culture.
View Article and Find Full Text PDFOral Versus Injectable Vitamin D Therapy for Treating Nutritional Rickets in Indian Children: A Comparative Study.
Indian J Orthop
February 2025
Department of Orthopaedics, JSS Medical College, Mysore, 570004 India.
Background: Rickets is a common metabolic bone disease in children, primarily caused by vitamin D deficiency. This study aimed to compare the efficacy of oral weekly vitamin D supplementation and injectable stoss therapy in treating nutritional rickets in Indian children.
Methods: This prospective, randomized, controlled trial was conducted over 18 months at a tertiary care center.
Line-Shifting Triarylmethyl Radicals for Imaging of Enzyme Activity Using Overhauser-Enhanced Magnetic Resonance Imaging: Application to Alkaline Phosphatase.
Chem Biomed Imaging
January 2025
In Vivo Multifunctional Magnetic Resonance Center, Robert C. Byrd Health Sciences Center, West Virginia University, Morgantown, West Virginia 26506, United States.
Enzyme catalytic activities are critical biomarkers of tissue states under physiological and pathophysiological conditions. However, the direct measurement and imaging of enzyme activity remains extremely challenging. We report the synthesis and characterization of the first stable triarylmethyl (TAM) radical substrate of alkaline phosphatase (TAM-ALPs).
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!