title: Lack of specificity shown by P2Y6 receptor antibodies.
abstract: P2Y6 receptor in bladder smooth muscle responds to UDP by increasing muscle tone and augmenting bladder contractions. The exact cellular location of the receptor is however unknown. Three commercially available antibodies to P2Y6 receptor gave clean bands on Western blot which were eliminated by specific peptide competition. Two of the three also immunostained bladder smooth muscle cells while leaving adjacent interstitial cells of Cajal unstained. However, attempts to validate the specificity of these antibodies by performing the same assays on bladders from P2Y6 knockout mice were unsuccessful. In Western blots, all three antibodies bound similar proteins in both wild type and P2Y6 knockout tissue. Immunostaining of knockout tissue sections also showed no difference in staining patterns or intensity. We conclude that rigorous controls are required when using commercial reagents to this G-protein coupled receptor and perhaps to other members of the P2Y receptor family.
authors.affiliation: Harvard Medical School, Harvard University, Boston, MA, USA.
authors.forename: Weiqun
authors.initials: W
authors.lastname: Yu
authors.forename: Warren G
authors.initials: WG
authors.lastname: Hill
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D066298:In Vitro Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009130:Muscle, Smooth / Q000378:metabolism*; D018048:Receptors, Purinergic P2 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D001743:Urinary Bladder / Q000378:metabolism*
chemical_list: D000906:Antibodies; D018048:Receptors, Purinergic P2; C099845:purinoceptor P2Y6