title: Immunoassay of in vitro activated human tissue transglutaminase.
abstract: Tissue transglutaminase (tTG) is a calcium-dependent enzyme that exerts a variety of physiological functions and is involved in various pathoprocesses. To characterize the role of tTG in disease, simple assays are necessary for detection. We developed a highly sensitive enzyme-linked immunosorbent assay (ELISA) that combines a transamidation step with immunological detection to determine tTG. This assay is based on covalent binding of in vitro activated tTG to N,N'-dimethylated casein and subsequent detection of coupled tTG by specific immunoglobulin G (IgG) antibodies directed against tTG followed by binding of a secondary biotin-labeled antibody. The assay reaches a detection limit of 0.05ng of tTG/ml. Type 1 and 3 transglutaminases and factor XIII are not detected. By use of this assay, tTG in several cell lines and the stimulatory effect of retinoic acid on tTG expression could be demonstrated. The new assay represents a promising tool for the study of tTG in normal cell physiology and under pathological conditions.
authors.affiliation: Institute of Laboratory Medicine, University Hospital Leipzig, D-04103 Leipzig, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Wolf
authors.forename: Ingolf
authors.initials: I
authors.lastname: Lachmann
authors.forename: Uta
authors.initials: U
authors.lastname: Wagner
authors.forename: Awad
authors.initials: A
authors.lastname: Osman
authors.forename: Thomas
authors.initials: T
authors.lastname: Mothes
mesh_terms: D000818:Animals; D001710:Biotin / Q000378:metabolism; D002103:Cadaverine / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D004789:Enzyme Activation / Q000187:drug effects; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007460:Iodoacetamide / Q000494:pharmacology; D051379:Mice; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D051381:Rats; D012015:Reference Standards; D011503:Transglutaminases / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology; D015032:Zinc / Q000494:pharmacology
chemical_list: D014212:Tretinoin; D001710:Biotin; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins; D015032:Zinc; D002103:Cadaverine; D002118:Calcium; D007460:Iodoacetamide