Electrophoretic detection of myeloperoxidase, protease, lactoferrin and lysozyme in buffalo polymorphonuclear granular acid extracts.

Polymorphonuclear (PMN) cells of more than 90% viability and 92% purity were isolated from the peripheral blood of buffaloes. The cationic proteins were extracted with 0.2 mol/L sodium acetate, pH 4.0 from the granules in the PMN and subjected to both non-denaturing and denaturing acid urea polyacrylamide gel electrophoresis (AUPAGE) for identification of myeloperoxidase (MPO), lysozyme, protease activity and lactoferrin. Protease was identified using alpha-naphthyl acetate as substrate, while lactoferrin was identified using a reference lactoferrin from bovine milk in AUPAGE, and by double immunodiffusion and Western blot techniques. Based on AUPAGE, lysozyme was found to be most cationic of all the proteins and peptides from the PMN granules as was evident from reference lysozyme run. The results indicated that the granules in buffalo PMN cells have lysozyme, protease, MPO and lactoferrin.