Analytical Behavior of Fura-2 and Its Determination of [Ca²⁺]_i in Lymphocytes Treated with Cefotaxime

【摘要】 The interaction of Fura-2 with Ca2+ is studied using steady fluorescence technique.The effect of pH on the spectra behavior of Fura-2 in the presence of Ca2+ 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-Ca2+ complex are examined,Kd is found to decrease with increasing temperatures(20 ℃,37 ℃,50 ℃)and ΔH is 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 Ca2+ concentration([Ca2+]i)in lymphocyte is developed by using Fura-2 as a fluorescence probe in the presence of Cefotaxime at 37 ℃ and pH 7.4.更多还原

【Abstract】 The interaction of Fura-2 with Ca2+ is studied using steady fluorescence technique.The effect of pH on the spectra behavior of Fura-2 in the presence of Ca2+ 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-Ca2+ complex are examined,Kd is found to decrease with increasing temperatures(20 ℃,37 ℃,50 ℃)and ΔH is 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 Ca2+ concentration([Ca2+]i)in lymphocyte is developed by using Fura-2 as a fluorescence probe in the presence of Cefotaxime at 37 ℃ and pH 7.4.更多还原