AMPA Receptor
Determining ad hoc experimental parameters for quantitative autoradiography of AMPAR using [3H]AMPA ligand
Experimental determination of receptor-ligand binding affinity for [3H]AMPA
10 nM [3H]AMPA bound 32% of receptors in rat. Frozen-thawed tissue in buffer with KCl bound 36% of receptors. Specific binding curve extrapolation estimates the Kd of AMPA to be ~28 nM (Olsen et al 1987).
| PROCEDURES | |||
|---|---|---|---|
| Radioligand | Pre-Incubation | Incubation | Wash |
| 3H-AMPA 15 nM |
50 mM Tris-HCl 100 mM KCl pH 7.4 10 min |
15 nM 3H-AMPA Pre-Incubation Buffer 60 min |
Pre-Incubation Buffer 10 min x2 baths 1 dip diH2O |