Investigation of PAR-1-type thrombin receptors in rat glioma C6 cells with a novel monoclonal anti-PAR-1 antibody (Mab COR7-6H9).
Rat glioma C6 cells have been demonstrated to be a suitable model in the investigation of PAR-1-type thrombin receptors in brain. However, anti-PAR-1 antibodies, which should be very helpful tools in studying PAR-1 in rat cells, have not been available up until now. Therefore, we prepared a monoclonal anti-thrombin receptor antibody (Mab COR7-6H9) directed against the peptide sequence GRAVYLNKSRFPPMPPPPFISEDASG in the N-terminus below the thrombin cleavage site of the rat PAR-1-type thrombin receptor. Using this antibody, we demonstrated the presence of PAR-1 binding sites on the plasma membrane of rat glioma C6 cells both with confocal laser fluorescence and with scanning electron microscopy. In addition, Mab COR7-6H9 was shown to block PAR-1-mediated transmembranal signaling as demonstrated by measurement of free intracellular calcium and cyclic AMP. This novel anti-PAR-1 antibody is therefore likely to be a very helpful tool in studying PAR-1-type thrombin receptors in rat brain.
Kaufmann
R
R
Research Group Pharmacological Hemostaseology Medical Faculty at the Friedrich-Schiller University Jena, Germany.
Bone
B
B
Westermann
M
M
Nowak
G
G
Ramakrishnan
V
V
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D005455:Fluorescent Antibody Technique; D005910:Glioma; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D051381:Rats; D044463:Receptor, PAR-1; D018179:Receptors, Thrombin / Q000276:immunology / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D044463:Receptor, PAR-1; D018179:Receptors, Thrombin