反应详情

The enhancement of calcium deposition between kaempferol and ipripfavone were compared. Test samples and their adding manner were same as Experiment 4. Calcium deposition amount was measured by usual method using “Calcium C Test Wako” (commercialized by Wako Pure Chemical Industries, Osaka, Japan); briefly, cultured cells were washed with 1 ml of Dulbecco phosphate buffer saline three times after removing the medium. To the resultant was added 0.5 ml of 2N hydrochloric acid to resolve deposited calcium. To 5 μl of the resolving solution were added 0.5 ml of 0.88M monoethanolamine buffer (pH11) and 0.05 ml of 0.63 mM o-cresolphthalein complexone solution (binding to calcium ion to form chelate complex) containing 69 mM 8-quinolinol and mixed, Calcium amount per one hole of the plate was calculated by measuring absorbance 570 nm of the resultants and standardizing with a standard calcium solution. The result of comparison with kaempferol and ipriflavone was shown in FIG. 2. The result of only ethanol treatment as a negative control is also shown in FIG. 2.

1. additionTest samples and their adding manner
2. washbriefly, cultured cells were washed with 1 ml of Dulbecco phosphate buffer saline three times
3. customafter removing the medium
4. additionTo the resultant was added 0.5 ml of 2N hydrochloric acid
5. additionTo 5 μl of the resolving solution were added 0.5 ml of 0.88M monoethanolamine buffer (pH11) and 0.05 ml of 0.63 mM o-cresolphthalein complexone solution (binding to calcium ion to form chelate complex)
6. additioncontaining 69 mM 8-quinolinol
7. additionmixed