Reaktionsgleichung
CCN[C:4]([C@H:6]1[O:10][C@@H:9]([N:11]2[C:15]3[N:16]=[CH:17][N:18]=[C:19]([NH2:20])[C:14]=3[N:13]=[CH:12]2)[C@H:8]([OH:21])[C@@H:7]1[OH:22])=[O:5].C1([C@@]2(N3C4N=CN=C(N)C=4N=C3)O[C@H](CO)[C@@H](O)[C@H]2O)CCCCC1.[Mg+2].[Cl-].[Cl-]>C1(N2C=NC3C2=NC=NC=3N[C@H](CC2C=CC=CC=2)CO)C=CC=CC=1>[C@@H:9]1([N:11]2[C:15]3[N:16]=[CH:17][N:18]=[C:19]([NH2:20])[C:14]=3[N:13]=[CH:12]2)[O:10][C@H:6]([CH2:4][OH:5])[C@@H:7]([OH:22])[C@H:8]1[OH:21]Komponenten
Edukt 3
Produkt 1
Reagenzien, Katalysatoren, Lösungsmittel 1
Versuchsvorschrift
Incubation tubes, in triplicate, receive 100 μl of [3H]NECA (94 nM in the assay), 100 μl of 1 μM cyclohexyladenosine (CHA), 100 μl of 100 mM MgCl2, 100 μl of 1 IU/ml adenosine deaminase, 100 μl of test compounds at various concentrations over the range of 10-10M to 10-4M diluted with assay buffer (50 mM Tris-HCl pH 7.7) and 0.2 μl of membrane suspension (5 mg wet weight), in a final volume of 1 ml of 50 mM Tris-HCl pH 7.7. Incubations are carried out at 25° C. for 60 minutes. Each tube is filtered through GF/B glass fiber filters using a vacuum. The filters are rinsed two times with 5 ml of the ice-cold buffer. The membranes on the filters are transferred to scintillation vials to which 8 ml of Omnifluor with 5% Protosol is added. The filters are counted by liquid scintillation spectrometry.
Aufarbeitung
- concentration/ml adenosine deaminase, 100 μl of test compounds at various concentrations over the range of 10-10M to 10-4M
- additiondiluted with assay buffer (50 mM Tris-HCl pH 7.7) and 0.2 μl of membrane suspension (5 mg wet weight)
- filtrationEach tube is filtered through GF/B glass fiber filters
- washThe filters are rinsed two times with 5 ml of the ice-cold buffer
- additionis added
Quellen und Nachweise
- ORD-Eintrag
- ord-6eaee0424e50499ca90416a9c2839a2e
- Quelldatensatz
- uspto-grants-1995_02
- Patentnummer
- US05391739
Verknüpfte Notizen
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