| Protocol | In Vitro | In vitro activity: AK enzyme inhibition was assayed radiochemically as described byYamada et al. (1980) and McNally et al. (1997). The ability of A-134974 to inhibit AK activity in intact IMR-32 neuroblastoma cells (American Type Culture Collection, Gaithersburg, MD) carried out as previously described (Kowaluk and Cowart, 1994). Radioligand binding assay methodology for the A1, A2A, and A3 receptors was carried out as described by Jarvis et al. (2000). The ability of A-134974 to inhibit [3H]nitrobenzylthioinosine binding to the ADO transporter and to inhibit adenosine deaminase activity was also examined using previously described methodology (Parkinson and Geiger, 1996). |
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| In Vivo | Drugs administered to rats were A-134974 (Cowart, 1997, an AK inhibitor synthesized at Abbott Laboratories), morphine sulfate (Mallinckrodt, St. Louis, MO), and theophylline (a nonselective ADO receptor antagonist; Sigma Chemical Co., St. Louis, MO). All drugs were dissolved in sterile water for local (intraplantar, i.t., or i.c.v.) or systemic (i.p.) delivery. The drug administration procedures described below were followed for locomotor activity experiments as well as for hyperalgesia experiments. Each experimental group consisted of at least five animals. |
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| Animal model | Male Sprague-Dawley rats (260–320 g) |
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These protocols are for reference only. InvivoChem does not
independently validate these methods.
Preparing Stock Solutions
| Solvent volume to be added |
Mass (the weight of a compound) |
| Mother liquor concentration |
1mg | 5mg | 10mg | 20mg |
| 1mM | 2.6655 mL | 13.3273 mL | 26.6546 mL | 53.3092 mL |
|---|
| 5mM | 0.5331 mL | 2.6655 mL | 5.3309 mL | 10.6618 mL |
|---|
| 10mM | 0.2665 mL | 1.3327 mL | 2.6655 mL | 5.3309 mL |
|---|
| 20mM | 0.1333 mL | 0.6664 mL | 1.3327 mL | 2.6655 mL |
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Quality Control Documentation
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