Mediators of Inflammation 
Volume 2006 (2006), Article ID 19307, 8 pages
doi:10.1155/MI/2006/19307
Research Communication

Application of Intracellular Alkaline Phosphatase Activity Measurement in Detection of Neutrophil Adherence In Vitro

Katarzyna Bednarska, Magdalena Klink, and Zofia Sulowska

 Department of Experimental Immunology, Centre for Medical Biology, Polish Academy of Sciences, Lodowa 106, Lodz 93-232, Poland
Received 28 April 2006; Accepted 13 July 2006

Abstract

We have proposed the use of the fluorimetric method with 4-methylumbelliferyl phosphate (4-MUP) specific substrate for the alkaline phosphatase determination in the neutrophil adhesion assay. We provide evidence that the endogenous neutrophil alkaline phosphatase (NAP) activity evaluation is reliable to quantify neutrophil adhesion at a wide range of cell numbers (104–106). The results obtained by fluorimetric NAP activity test correlate to the results of adherence evaluated using the MTT reduction assay. The fluorimetric NAP activity test may be applied for resting as well as activated neutrophils without the risk of the activators interferences into the test. The alkaline phosphatase survey with the use of 4-MUP substrate is recommended herein as a sensitive, repeatable, simple, and reliable method of the neutrophil adherence determination in vitro.