Detection of human and rodent 5-HT3B receptor subunits by anti-peptide polyclonal antibodies
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Reeves, D., & Lummis, S. (2006). Detection of human and rodent 5-HT3B receptor subunits by anti-peptide polyclonal antibodies. BMC NEUROSCIENCE, 7 https://doi.org/10.1186/1471-2202-7-27
Background: The 5-HT3 receptor is a member of a neurotransmitter-gated ion channel family which includes nicotinic acetylcholine, GABA(A), and glycine receptors. While antibodies specific for the 5-HT3A receptor subunit are plentiful, and have revealed a wealth of structural and functional information, few antisera exist for the detection of 5-HT3B receptor subunits. Here we describe the generation and characterisation of a rabbit polyclonal antiserum that specifically recognises 5HT(3B) receptor subunits|Results: Immunization of a rabbit with a 20-mer peptide, corresponding to the N-terminus of the human 5-HT3B receptor subunit, generated serum with polyclonal antibodies from which an IgG fraction was purified, yielding pAb77. The antibodies were shown to label 5-HT3B receptor subunits in transfected human embryonic kidney cells and rodent tissues using Western blots. Immunocytochemistry using pAb77 on these cells showed that 5-HT3B receptor subunits do not reach the plasma membrane in the absence of 5-HT3A receptor subunits. Immunohistochemical analysis of rat brain sections showed pAb77 immunoreactivity in distinct populations of cells in the hippocampus.|Conclusion: We have demonstrated that pAb77 antibodies specifically label native and recombinant 5-HT3B receptor subunits with high affinity and specificity. The antibody was shown to be useful for the determination of both receptor trafficking and also mapping 5-HT3B receptor subunit expression in the CNS.
neurons, CNS, NICOTINIC ACETYLCHOLINE-RECEPTOR, GATED ION-CHANNEL, 5-HYDROXYTRYPTAMINE TYPE-3 RECEPTORS, ALTERS, CELL-SURFACE EXPRESSION, SEROTONIN-RECEPTOR
External DOI: https://doi.org/10.1186/1471-2202-7-27
This record's URL: http://www.dspace.cam.ac.uk/handle/1810/238071
Rights Holder: Reeves et al.; licensee BioMed Central Ltd.