Reaktion #6781

Reaktionsgleichung

Tris HCl

MgCl2

[3H]PGD2

→

PGD2

Reaktionsbedingungen

Aufarbeitung

1. 1
   Sonstigethe mixture was reacted at 4° C. for 90 min
2. 2
   SonstigeAfter the reaction
3. 3
   Filtrationthe mixture was filtered through a glass fiber
4. 4
   Filtrationfilter paper
5. 5
   Waschenwashed several times
6. 6
   Temperaturwith cooled physiological saline
7. 7
   EinengenThe inhibitory activity of each compound was expressed as the concentration

Vorschrift

To a binding-reaction solution (50 mM Tris/HCl, pH 7.4, 5 mM MgCl2) (0.2 ml) were added the human platelet membrane fraction (0.1 mg) and 5 nM [3H]PGD2 (115 Ci/mmol), and the mixture was reacted at 4° C. for 90 min. After the reaction, the mixture was filtered through a glass fiber filter paper and washed several times with cooled physiological saline, then the radioactivity retained on the filter paper was measured. The specific-binding ratio was calculated by subtracting the non-specific binding ratio which is the radioactivity similarly measured in the presence of 10 μM PGD2 from the total binding. The inhibitory activity of each compound was expressed as the concentration required for 50% inhibition (IC50), which was determined by depicting a substitution curve by plotting the binding ratio (%) in the presence of each compound, where the binding ratio in the absence of a test compound is 100%.

Quelle