Pierce chemiluminescent substrates, solution A and solution B, were mixed at a ratio of 1 1 for coloration at room temperature for 5 minutes, then the reaction mixture was aspirated, and the fluorescence/chemiluminescence imaging system (Fuji Film, Tokyo, Japan) was used for analysis.
After confirmation of clear cerebrospinal fluid (CSF) efflux, solution A (n =60) or solution B (n =60) was administered.
At the start of the treatment the solutions in both tanks were replaced with either solution A, identical to that above, or solution B, identical to that above except that the KH2PO4 content was reduced to 0·016 m m.
Figure 1 NaOH (or sample solution) (a), CdTe solution (b), NaClO solution (c), and H 2 O 2 solution (d). Figure 2 CL kinetic curves of H 2 O 2 -CdTe NC CL reaction.
Neurons were voltage-clamped at +40 mV (intrapipette solution A) or at −70 mV (intrapipette solution B) holding potential, using an Axopatch 200B amplifier (Axon Instruments, Foster City, CA).
Aqueous copper acetate and aqueous NaOH solutions were referred to as solution A and solution B, respectively.
First, the precursor was prepared by mixing solution A with solution B (1.5 100 in v/v).
For a typical synaptic NMDAR stimulation experiment, crude synaptosomes (∼400 µg) were pre-incubated with 100 µM glycine for 20 min. at 37°C in solution B, followed by the addition of either solution B alone (control) or solution B containing high KCl (50 mM final concentration) for various times (5 to 120 min).
After successive washing with solution A and solution B (0.1 % TFA and 80%% acetonitrile), the resulting phosphopeptides were eluted by 0.5 and 5 % piperidine (Wako).
Label this beaker as "catalyst solution" or "solution B".
