Reaktionsgleichung
C1N(CCO)CCN(CCS(O)(=O)=O)C1.N[C@H:17]([C:23]([OH:25])=[O:24])[CH2:18][CH2:19][C:20](=O)N.[CH3:26][C:27]1(C)S[C@@H]2[C@H](NC(CC3C=CC=CC=3)=O)C(=O)[N:29]2[C@H:28]1[C:45]([O-])=O.[K+].[CH3:50][C@@H:51]1O[C@@H:54]([O:56][C@H:57]2[C@H:62](O)[C@@H:61](O)[C@H:60](NC(N)=N)[C@@H:59]([OH:69])[C@@H:58]2NC(N)=N)[C@H:53](O[C@@H]2O[C@@H](CO)[C@H](O)[C@@H](O)[C@@H]2NC)[C@@:52]1([OH:89])C=O.C[S:91]([CH3:93])=O>>[CH:27]1[C:28]([N:29]=[C:93]=[S:91])=[CH:45][C:17]2[C:23]([O:25][C:19]3([C:20]4[CH:50]=[CH:51][C:52]([OH:89])=[CH:53][C:54]=4[O:56][C:57]4[CH:58]=[C:59]([OH:69])[CH:60]=[CH:61][C:62]3=4)[C:18]=2[CH:26]=1)=[O:24]Komponenten
Versuchsvorschrift
Jurkat cells were grown in RPMI 1640 media containing 25 mM Hepes and L-glutamine (Gibco) supplemented with 10% FCS and penicillin/streptomycin and harvested by centrifugation (200×g, 10 minutes) when they reached a concentration of 1×106 cells/mL. 1×109 Jurkat cells were resuspended in 100 mL RPMI 1640 media with 0.5% FCS. Cells were then treated with either 1% DMSO or 5 μM 5-[(5-gambogylaminopentyl)-thioureidyl]-fluorescein (Example 10) in DMSO for 30 minutes at 37° C. Cells were washed one time with 40 mL phosphate buffered saline (PBS) and lysed in 8 mL RIPA buffer (10×RIPA supplied by Upstate) and 0.1% Protease Inhibitor Cocktail (Sigma). The lysed cells were spun at 20,000×g for 10 minutes and the supernatant collected and referred to as “Jurkat lysate.”
Aufarbeitung
- customharvested by centrifugation (200×g, 10 minutes) when they
- washCells were washed one time with 40 mL phosphate buffered saline (PBS)
- customthe supernatant collected
Quellen und Nachweise
- ORD-Eintrag
- ord-57faede37ca44eedb0e7264a8f9899a0
- Quelldatensatz
- uspto-grants-2009_09
- Patentnummer
- US07592143B2
Verknüpfte Notizen
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