用自制新型荧光剂EPQS标记蛋白质及其荧光纸层析扫描测定

The Qualitative and Quantitative Analyses of Protein with a Novel Fluorescence Agent (EPQS) by Fluorescent Scanning on Solid Substrate

用自行研制的新型水溶性荧光剂EPQS分别标记人血清白蛋白(HSA)、牛血清白蛋白(BSA)和猪胰岛素(Insulin),经乙酸纤维素薄膜电泳纯化后,作荧光纸层析扫描,测得EPQS-HSA、EPQS-BSA和EPQS-Insulin的荧光光谱(λ_(em))分别为470、465、468nm,激发光谱(λ_(ex))分别为370、373、371nm;EPQS的λ_(em)为476nm,λ_(ex)为382nm。表明EPQS能用于蛋白质的标记。用EPQS定量HSA,结果显示,1～5μg的EPQS-HSA作荧光纸层析扫描,其结果与荧光强度有良好的线性关系。

We synthezised a newfluorescence agent, EPQS, which could besoluble in water and was stable in theanalytical process. It was characteristic ofgood fluorescence parameters. Humanserum albumin (HSA) was marked withEPQS and purified by the paper eletro-phoresis dialysis. We determined the fluo-rescent spectra and excitation spectra ofthe marked HSA, or EPQS on the cellu-lose acetate paper by RF-510 fluorescencespectrophotometer-thin-layer chromatos-canner. The formerλex=370nmλem=47nm;the latterλex= 382nmλem=476nm. Then wetouched different quantity of EPQS-HSAupon the cellulose acetate. The fluorescentscaning on the paper disclosed that therelationship between the quantity adoptedand the fluorescence intensity is linear.These showed that the protein could bequickly determined qualitatively and qu-antitatively by this simple method, andEPQS was useful.