Polyclonal antibodies against malondialdehyde-modified proteins: characterization and application in study of in vitro lipid peroxidation of cellular membranes.
In order to develop an immunofluorescent technique able to detect the cellular damage induced by lipid peroxidation, we produced rabbit polyclonal antibodies (AbAIP) against malondialdehyde (MDA)-modified lysozyme. These antibodies specifically recognized all the MDA-treated proteins tested, but not native proteins or proteins treated by other aldehydes. Moreover, we showed by an enzyme-linked immunosorbent assay (ELISA) that the amount of immunoreactive protein in MDA-treated erythrocytes was proportional to the concentration of MDA added, suggesting that this ELISA measurement may represent a quantitative determination of peroxidative protein alterations. In addition, when coupled to an indirect fluorochrome antibody conjugate [fluorescein isothiocyanate (FITC)], these antibodies permitted precise localization of the modified proteins within the membranes of peroxidized cells.
Petit
E
E
Laboratoire de Neurosciences, URA 1829 CNRS, Caen, France.
Chancerelle
Y
Y
Dumont
E
E
Divoux
D
D
Kergonou
F
F
Nouvelot
A
A
D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000378:metabolism; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004910:Erythrocyte Membrane / Q000276:immunology / Q000378:metabolism* / Q000528:radiation effects; D006801:Humans; D015227:Lipid Peroxidation / Q000502:physiology*; D008315:Malondialdehyde / Q000494:pharmacology*; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009113:Muramidase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D018384:Oxidative Stress
D000906:Antibodies; D008565:Membrane Proteins; D008315:Malondialdehyde; D009113:Muramidase