unlabeled α bgtx  (Alomone Labs)

Journal: Science Advances

Article Title: Electrically silent mutants unravel the mechanism of binding−gating coupling in Cys-loop receptors

doi: 10.1126/sciadv.adq8048

Figure Lengend Snippet: ( A ) Nicotine. ( B ) Carbamylcholine. ( C ) TMA. ( D ) MLA. The curves were fitted with single-component Hill equations (table S1), and for all of them, the labeled ligand was [ 125 I]-α-BgTx at a concentration (of the unbound form) of ~1 × K D,closed (fig. S3). The color code is the same for all panels.

Article Snippet: [I]-α-BgTx was purchased from Revvity, unlabeled α-BgTx from Alomone Labs, MLA and PNU-120596 from Tocris Bioscience, and ACh, carbamylcholine, nicotine, TMA, IVM, etomidate, propofol, picrotoxinin, lindane, and menthol, from MilliporeSigma.

Techniques: Labeling, Concentration Assay

Journal: Science Advances

Article Title: Electrically silent mutants unravel the mechanism of binding−gating coupling in Cys-loop receptors

doi: 10.1126/sciadv.adq8048

Figure Lengend Snippet: ( A to F ) α-BgTx–nicotine curves of the indicated constructs. The insertion of one or two residues in the pre-M1 linker markedly reduced the rightward shift caused by IVM. ( G ) α-BgTx–MLA curves of the wild-type construct. The curves were fitted with single-component Hill equations (table S1), and for all of them, the labeled ligand was [ 125 I]-α-BgTx at a concentration (of the unbound form) of ~1 × K D,closed (fig. S3).

Article Snippet: [I]-α-BgTx was purchased from Revvity, unlabeled α-BgTx from Alomone Labs, MLA and PNU-120596 from Tocris Bioscience, and ACh, carbamylcholine, nicotine, TMA, IVM, etomidate, propofol, picrotoxinin, lindane, and menthol, from MilliporeSigma.

Techniques: Construct, Labeling, Concentration Assay

Journal: Science Advances

Article Title: Electrically silent mutants unravel the mechanism of binding−gating coupling in Cys-loop receptors

doi: 10.1126/sciadv.adq8048

Figure Lengend Snippet: ( A ) Lack of effect of residue insertions in the pre-M1 linker on the affinity of the channel’s closed state for MLA. ( B to E ) Effect of menthol on the α-BgTx–nicotine competition curves. The insertion of residues in the pre-M1 linker reduced the shift caused by menthol. ( F ) Lack of effect of menthol on the affinity of the wild-type’s closed state for MLA. The curves were fitted with single-component Hill equations (table S1), and for all of them, the labeled ligand was [ 125 I]-α-BgTx at a concentration (of the unbound form) of ~1 × K D,closed (fig. S3). Unlike the competition curves obtained with the α7-AChR–β-GluCl chimera, some of the curves obtained with the full-length α7-AChR required Hill coefficients that were lower than unity to be best-fitted. Although different phenomena can underlie this finding , we tentatively favor the idea that this construct may approach equilibrium more slowly.

Article Snippet: [I]-α-BgTx was purchased from Revvity, unlabeled α-BgTx from Alomone Labs, MLA and PNU-120596 from Tocris Bioscience, and ACh, carbamylcholine, nicotine, TMA, IVM, etomidate, propofol, picrotoxinin, lindane, and menthol, from MilliporeSigma.

Techniques: Residue, Labeling, Concentration Assay, Construct