Application of intracellular alkaline phosphatase activity measurement in detection of neutrophil adherence in vitro.
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 (10(4)-10(6)). 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.
Bednarska
Katarzyna
K
Department of Experimental Immunology, Centre for Medical Biology, Polish Academy of Sciences, Ladowa, Lodz, Poland.
Klink
Magdalena
M
Sulowska
Zofia
Z
D000469:Alkaline Phosphatase / Q000378:metabolism*; D002448:Cell Adhesion; D005470:Fluorometry / Q000379:methods; D006801:Humans; D006923:Hymecromone / Q000031:analogs & derivatives / Q000378:metabolism; D009504:Neutrophils / Q000166:cytology* / Q000201:enzymology; D013379:Substrate Specificity
C005359:4-methylumbelliferyl phosphate; D006923:Hymecromone; D000469:Alkaline Phosphatase