Tissue inhibitor of metalloproteases (TIMP) is matrix associated in aortic tissue: report of a radioimmunoassay.
Tissue inhibitor of metalloproteinases (TIMP) is the major inhibitor of collagenase, gelatinase, proteoglycanase, stromelysin, and metalloelastases. An imbalance between proteases and inhibitors has been implicated in numerous disease processes including tumor invasion, rheumatoid arthritis, emphysema, and aortic aneurysm disease. The purpose of this investigation was to develop a polyclonal antibody to recombinant TIMP and establish an immunoassay to measure immunoreactive protein in normal and diseased tissues. A polyclonal antibody was produced in rabbit against recombinant human TIMP which was characterized and used to establish a radioimmunoassay. The assay was used to measure immunoreactive protein in fibroblast conditioned medium, human serum, and aortic extracts. There was more immunoreactive TIMP in matrix associated urea extracts than soluble salt extracts from human aorta, suggesting that TIMP is matrix associated. The sensitivity of the assay enables the specific measurement of this inhibitor in serum, fibroblast culture medium, and tissue extracts.
Brophy
C M
CM
Department of Surgery, Yale University School of Medicine, New Haven, CT 06510.
Marks
W H
WH
Reilly
J M
JM
Tilson
M D
MD
D001012:Aorta, Abdominal / Q000032:analysis*; D002478:Cells, Cultured; D005347:Fibroblasts / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis* / Q000097:blood / Q000276:immunology; D006801:Humans; D007106:Immune Sera; D007122:Immunoelectrophoresis; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors*; D009131:Muscle, Smooth, Vascular / Q000032:analysis*; D011863:Radioimmunoassay / Q000379:methods; D011994:Recombinant Proteins / Q000032:analysis; D012867:Skin / Q000032:analysis; D019714:Tissue Inhibitor of Metalloproteinases
D006023:Glycoproteins; D007106:Immune Sera; D011994:Recombinant Proteins; D019714:Tissue Inhibitor of Metalloproteinases; D008666:Metalloendopeptidases