ELECTROPHORETIC DETECTION OF MYELOPEROXIDASE, PROTEASE, LACTOFERRIN AND LYSOZYME IN BUFFALO POLYMORPHONUCLEAR GRANULAR ACID-EXTRACTS

Polymorphonuclear (PMN) cells of more than 90% viability and 92% purit y were isolated from the peripheral blood of buffaloes. The cationic p roteins 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 denaturin g acid urea polyacrylamide gel electrophoresis (AUPAGE) for identifica tion of myeloperoxidase (MPO), lysozyme, protease activity and lactofe rrin. Protease was identified using alpha-naphthyl acetate as substrat e, 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 o f all the proteins and peptides from the PMN granules as was evident f rom reference lysozyme run. The results indicated that the granules in buffalo PMN cells have lysozyme, protease, MPO and lactoferrin.