6533b837fe1ef96bd12a3418

RESEARCH PRODUCT

Identification and characterization of a monoclonal antibody to the membrane fatty acid binding protein

Wolfgang StremmelJohannes GunawanMichael MannsHans Erich DiedeEnrique Rodilla

subject

medicine.drug_classBlotting WesternImmunoblottingBiophysicsFluorescent Antibody TechniqueEnzyme-Linked Immunosorbent AssayNerve Tissue ProteinsFatty Acid-Binding ProteinsMonoclonal antibodyBiochemistryFatty acid-binding proteinCell LineMiceEndocrinologyAffinity chromatographymedicineAnimalsHumanschemistry.chemical_classificationMice Inbred BALB CbiologyMembrane transport proteinTumor Suppressor ProteinsBinding proteinCell MembraneFatty AcidsAntibodies MonoclonalFatty acidMolecular biologyNeoplasm ProteinsRatsLiverchemistryMembrane proteinBiochemistrybiology.proteinElectrophoresis Polyacrylamide GelAntibodyCarrier ProteinsFatty Acid-Binding Protein 7

description

A monoclonal antibody to the rat liver membrane fatty acid binding protein (MFABP) was prepared by immunizing mice with purified MFABP isolated from solubilized rat liver plasma membrane proteins by oleate-agarose affinity chromatography technique. The monoclonal antibody K15/6 identified a single 40 kDa protein in rat liver plasma membranes with pI values of 8.5, 8.8 and 9.0, which is identical to the authentic MFABP, but clearly distinct from rat mitochondrial GOT. The antibody K15/6 selectively inhibited cellular influx as well as membrane binding of fatty acids, but not of cholesterol or vitamin E. The same antibody was used in immunofluorescence, ELISA and Western blot analysis to determine the subcellular and organ distribution pattern of MFABP. The protein was identified in rat liver plasma membranes and mitochondria, but in no other cell compartment. It was detectable in homogenates of rat liver but not in homogenates of other organs. Therefore, the monoclonal antibody K15/6 represents an organ specific antibody to MFABP which reveals inhibitory action on membrane binding/transport of fatty acids.

https://doi.org/10.1016/0005-2760(92)90149-p