Isolation and characterization of myeloperoxidase from leukocytes of rat peritoneal fluid.
Myeloperoxidase (MPO) was isolated from rat peritoneal leukocytes with a yield of 51% and A430/A280 = 0.75 - 0.80, and its physicochemical properties were studied. The molecular weight of the MPO is about 150 kD. The MPO was assayed for amino acid content. We used substrate mixture containing phenol, 4-aminoantipyrine, and H2O2 to detect 10(-10) M of the enzyme. The MPO was localized in rat blood neutrophils using polyclonal anti-MPO antibodies and secondary fluorescein isothiocyanate-labeled antibodies. Immunofluorimetric assay (IFMA) was developed for quantitative measurement of the MPO. The MPO and leukocytes can iodinate BSA using NaI or thyroxine as the source of iodine.
Morozov
V I
VI
Department of Biochemistry, Institute of Physical Culture, St. Petersburg, Russia.
Tsyplenkov
P V
PV
Kokryakov
V N
VN
Volkov
K N
KN
Vinogradova
N A
NA
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D001202:Ascitic Fluid / Q000166:cytology* / Q000201:enzymology*; D002417:Cattle; D005456:Fluorescent Dyes; D006219:Halogens; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007962:Leukocytes / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D009504:Neutrophils / Q000201:enzymology; D009195:Peroxidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D012710:Serum Albumin, Bovine; D013379:Substrate Specificity
D000596:Amino Acids; D000906:Antibodies; D005456:Fluorescent Dyes; D006219:Halogens; D012710:Serum Albumin, Bovine; D009195:Peroxidase