鈣黃綠素乙酰甲酯性質(zhì)、用途與生產(chǎn)工藝
鈣黃綠素AM是一種膜可滲透的非熒光分子。它可通過(guò)細(xì)胞內(nèi)酯酶轉(zhuǎn)化為熒光鈣黃綠素。鈣黃綠素具有有限的膜滲透性。鈣黃綠素AM也是p-糖蛋白(Pgp)的底物。Pgp的存在會(huì)導(dǎo)致鈣黃綠素AM的流出，從而降低胞內(nèi)的熒光。另一方面，Pgp抑制劑的存在可增加鈣黃綠素AM的細(xì)胞進(jìn)入，從而增加細(xì)胞的熒光。鈣黃綠素AM在早期藥物發(fā)現(xiàn)中起著重要作用。鈣黃綠素的熒光細(xì)胞可滲透衍生物。
Calcein-AM是用于測(cè)定細(xì)胞活力的可以滲透細(xì)胞的熒光染料。
The calcein-AM dye used to stain the living cells is shown to have a low spontaneousleakage rate less than 15% in 4 hours at 37°C. Dilutions of targets stained by calcein-AM has a linear relationship with measured fluorescence values. NK cells, LAKs, and CTLs are readily detectable by this microtest. Quantitation of killing and kinetic analysis is readily performed with the test system. Calcein-AM is pH independent, better retained and more photostable. In addition, the high level of intracellular retention of calcein-AM and its low-level release after incorporation exclude possible cell-monolayer labeling and allow its use in a cell-cell interaction assay. Moreover, the bright fluorescence can easily be detected and measured by a microplate fluorescence reader. Calcein-AM is a highly lipophilic vital dye that rapidly enters viable cells, is converted by intracellular esterases to calcein that produces an intense green (530-nm) signal, and is retained by cells with intact plasma membrane. From dying or damaged cells with compromised membrane integrity or from cells expressing multidrug resistance protein (MRP), unhydrolyzed substrates and their fluorescent products are rapidly extruded from cells. The calcein-AM assay has been used to assess the cell viability, cytotoxicity and tp quantitate apoptosis.
Calcein-AM is found to be suitable for
in vivo
studies, because it has no deleterious effects on cell function and is, indeed, a marker of cell viability.
用途
鈣黃綠素-AM 細(xì)胞可滲透化合物常被用做絡(luò)合物指示劑，用來(lái)滴定EDTA螯合的鈣離子，并且用熒光檢測(cè)法測(cè)定鈣離子濃度。它具有非常好的水溶性，其質(zhì)子解離常數(shù)（酸度系數(shù)）pKa1=2.1, pKa2=2.9, pKa3=4.2, pKa4=5.5, pKa5=10.8和pKa6=11.7。
Calcein-AM是一種可對(duì)活細(xì)胞進(jìn)行熒光標(biāo)記的細(xì)胞染色試劑，Calcein-AM由于在Calcein(鈣黃綠素)的基礎(chǔ)上加強(qiáng)了疏水性，因此能夠輕易穿透活細(xì)胞膜。當(dāng)其進(jìn)入到細(xì)胞質(zhì)后，酯酶會(huì)將其水解為Calcein(鈣黃綠素)留在細(xì)胞內(nèi),發(fā)出強(qiáng)綠色熒光。與其它同類試劑（如BCECF-AM和Carboxy-fluorescein diacetate）相比，Calcein-AM是最適合作為熒光探針去染活細(xì)胞的，因?yàn)樗募?xì)胞毒性很低。
鈣黃綠素乙酰甲酯
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