A reference method for measurement of alkaline phosphatase activity in human serum.

Abstract

We present an official AACC reference method for the measurement of alkaline phosphatase, the culmination of optimization experiments conducted by a group of independent laboratories. The details of this method and evaluation of factors affecting the measurement are described. A metal ion buffer has been incorporated that maintains optimal and constant concentrations of zinc(II) and magnesium(II) ions. Final reaction conditions are: pH (30 degrees C), 10.40 +/- 0.05; 2-amino-2-methyl-1-propanol buffer, 0.35 mol/L; 4-nitrophenyl phosphate, 16.0 mmol/L; magnesium acetate, 2.0 mmol/L; zinc sulfate, 1.0 mmol/L; and N-(2-hydroxyethyl)ethylenediaminetriacetic acid, 2.0 mmol/L.

Stimulative Effect of Ginsenosides Rg5:Rk1 on Murine Osteoblastic MC3T3‐E1 Cells

“…Protein contents were analyzed using a Smart BCA protein assay kit (iNtRON Biotechnology, Korea) according to the manufacturer's recommended protocol. ALP activities were determined by adding the p-nitrophenyl phosphate as substrate at 37°C for 1 h. Reactions were then stopped by the addition of 50 μL 1 N NaOH (Tietz et al, 1983). The absorbances were then measured at 405 nm using a microplate reader.…”

Human periodontal fibroblasts viability stored in Custodiol®, coconut water, and propolis. An ex vivo study

“…Then the rate of increase in absorbance at 405 nm was recorded for 5 minutes. [23][24][25] One day before treatment, cells were seeded in 96-well tissue culture plates (Nunk, Denmark) by trypsinization of cells in the culture flasks, counting them using trypan blue, and seeding them with the concentration of 16 000 cells/well. After that, the 96-well plates were incubated at 37°C in a humidified atmosphere of 5% CO 2 in air.…”

Ginsenoside Rh1 induces mouse osteoblast growth and differentiation through the bone morphogenetic protein 2/runt-related gene 2 signalling pathway

“…The reaction was stopped by adding 1-m NaOH and the absorbance was measured at 405 nm by an ELISA reader. [34] Protein concentration was determined by incubation in SMART micro BCA Protein Assay Kit (Intron Biotechnology, Inc., Gyeonggi-do, Korea) according to manufactured protocol.…”