Detection of the human GPR50 orphan seven transmembrane protein by polyclonal antibodies mapping different epitopes.

Abstract:

:GPR50 is an orphan seven transmembrane protein related to the melatonin receptor subfamily comprising MT(1) and MT(2) receptors. In the absence of any known ligand for GPR50, other tools are critical for the characterization of this protein. Here, we describe the generation, purification and characterization of the first rabbit polyclonal antibodies generated against peptides corresponding to the N-terminus, C-terminus and two additional regions within the intracellular tail of GPR50. Immune sera were purified on peptide-antigen affinity columns. Antibodies specifically recognized a GPR50-YFP fusion protein on the plasma membrane of HEK 293 cells in immunofluorescence experiments. In Western blot experiments, the monomeric and dimeric forms of GPR50 were detected as proteins of 66 and 130 kDa, respectively. In addition, these new antibodies were sufficiently sensitive to detect GPR50 in brain slices of the rat pituitary and human hippocampus. In conclusion, we successfully produced antibodies against the orphan GPR50 protein that will become valuable tools for functional studies of this protein.

journal_name

J Pineal Res

authors

Hamouda HO,Chen P,Levoye A,Sözer-Topçular N,Daulat AM,Guillaume JL,Ravid R,Savaskan E,Ferry G,Boutin JA,Delagrange P,Jockers R,Maurice P

doi

10.1111/j.1600-079X.2007.00437.x

subject

Has Abstract

pub_date

2007-08-01 00:00:00

pages

10-5

issue

1

eissn

0742-3098

issn

1600-079X

pii

JPI437

journal_volume

43

pub_type

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