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메나디온에 의한 혈소판 내 칼슘 변화측정시 형광 색소 사용의 문제점
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  • 메나디온에 의한 혈소판 내 칼슘 변화측정시 형광 색소 사용의 문제점
  • Infeasibility of Measuring $Ca^{2+}$ in Menadione-Exposed Platelets Using Fluorescent Dyes
저자명
정선화,이무열,이주영,정승민,정진호,Chung. Sun-Hwa,Lee. Moo-Yeol,Lee. Joo-Young,Chung. Seung-Min,Chung. Jin-Ho
간행물명
약학회지
권/호정보
1997년|41권 6호|pp.749-755 (7 pages)
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대한약학회
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정기간행물|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

It has been reported that dose-dependent $Ca^{2+}$ increase by menadione in platelets could be measured by fluorescent dye, quin-2. The problems will be described here rel ating to measuring $Ca^{2+}$ in menadione-exposed platelets using fura-2 and fluo-3, widely used fluorescent indicators. Additions of menadione to fura-2 loaded platelets and their lysates resulted in marked reduction in fluorescence intensity at both 340nm ($Ca^{2+}$-unbound form) 380nm ($Ca^{2+}$-undbound form) excitation wavelengths. Fura-2 excitation spectra were overlapped with UV-visible absorption spectra of menadione, suggesting that light absorption by menadione itself could quench fluorescence generated by fura-2. Next approach was to use fluo-3 which has the higher wavelength (490nm) of excitation. Previous work demonstrated that treatment with probenecid to platelets was required to prevent fluo-3 dye leakage. However, probenecid itself was proven to be inadequate to measure the concentration of intracellular $Ca^{2+}$; by reducing menadione-induced cytotoxicity in platelets. Our results suggest that it is not feasible to measure $Ca^{2+}$ in platelets by using fura-2 and fluo-3 in the presence of probenecid, and cautions should be taken to measure changes of intracellular $Ca^{2+}$ levels by fluorescent dyes following chemical exposure.