The interaction of Fura-2 with Ca^2+ is studied using steady fluorescence technique. The effect of pH on the spectra behavior of Fura-2 in the presence of Ca^2+ is investigated, the excitation maxima (340 nm) and ...The interaction of Fura-2 with Ca^2+ is studied using steady fluorescence technique. The effect of pH on the spectra behavior of Fura-2 in the presence of Ca^2+ is investigated, the excitation maxima (340 nm) and the isobestic point (360 nm) for the fluorescence spectra of Fura-2 depend on pH. At different temperatures the apparent dissociation constants ( Kd ) of Fura-2-Ca^2+ complex are examined, Kd is found to decrease with increasing temperatures (20 ℃, 37 ℃, 50 ℃ ) and △His calculated to be 21.16 kJ/mol by using the Van' t Hoff equation at pH 7.4 for all the temperatures tested. The determination of intracellular Ca^2+ concentration ( [Ca^2+ ] i ) in lymphocyte is developed by using Fura-2 as a fluorescence probe in the presence of Cefotaxime at 37 ℃ and pH 7.4.展开更多
基金supported by the Key Project of National Nature Science Foundation of China(NO.90813018)the Returned Chinese Scholars Fund
文摘The interaction of Fura-2 with Ca^2+ is studied using steady fluorescence technique. The effect of pH on the spectra behavior of Fura-2 in the presence of Ca^2+ is investigated, the excitation maxima (340 nm) and the isobestic point (360 nm) for the fluorescence spectra of Fura-2 depend on pH. At different temperatures the apparent dissociation constants ( Kd ) of Fura-2-Ca^2+ complex are examined, Kd is found to decrease with increasing temperatures (20 ℃, 37 ℃, 50 ℃ ) and △His calculated to be 21.16 kJ/mol by using the Van' t Hoff equation at pH 7.4 for all the temperatures tested. The determination of intracellular Ca^2+ concentration ( [Ca^2+ ] i ) in lymphocyte is developed by using Fura-2 as a fluorescence probe in the presence of Cefotaxime at 37 ℃ and pH 7.4.