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. 2017 Sep 4;8:77. doi: 10.3389/fpsyt.2017.00077

Table 1.

Summary of autoradiographic binding methods.

Radioligand (target)	Preincubation	Specific binding conditions	Non-specific binding condition	Wash
[³H]MK-801 (42) (N-methyl-d-aspartate receptor channel)		2.5 h at RT	20 µM non-tritiated MK-801 (Sigma)	2 × 20 min at 4°C
[³H]MK-801 (42) (N-methyl-d-aspartate receptor channel)		30 mM HEPES buffer with 100 µM glycine, 100 µM glutamate, 1 mM EDTA, and 20 nM [³H]MK-801 (s.a. 17.1 Ci/mmol, PerkinElmer, USA), pH 7.5	20 µM non-tritiated MK-801 (Sigma)	30 mM HEPES buffer with 1 mM EDTA, pH 7.5
[³H]CGP39653 (43) (NR2A subunit)	45 min at RT	45 min at RT	1 mM l-glutamic acid	30 s at 4°C
[³H]CGP39653 (43) (NR2A subunit)	50 mM Tris–HCl, pH 8	50 mM Tris–HCl with 20 nM [³H]CGP39653, pH 8	1 mM l-glutamic acid	50 mM Tris–HCl, pH 8
[³H]Ifenprodil (NRB subunit)		3 h at 4°C	10 µM non-tritiated Ifenprodil	3 × 5 min at 4°C
[³H]Ifenprodil (NRB subunit)		50 mM Tris–HCI buffer with 3 μM R(+)-3-(3-hydroxyphenyl)-N-propylpiperidine hydrochloride, 30 µM GBR-12909, 100 µM 20 nM [³H]Ifenprodil trifluoperazine, pH 7.4	10 µM non-tritiated Ifenprodil	50 mM Tris–HCl buffer, pH 7.4