CAJ | 학술논문

目的：建立一种简单、快速、高通量的检测GFP蛋白的方法。方法：用TECAN多功能酶标仪Infinite M200pro检测GFP蛋白在不同缓冲液中的荧光强度；用酶标仪检测含GFP的蛋白样品梯度稀释后的荧光强度，建立荧光强度与GFP相对含量之间的标准曲线；分别比较荧光酶标仪的检测结果与流式细胞术和Western-blotting检测结果的相关性。结果：使用50 mmol/L Tris-HCl(pH=10)的缓冲液稀释GFP蛋白更有利于检测低浓度的GFP；荧光酶标仪检测GFP的荧光读数同GFP相对含量之间的回归系数2R>0.99。荧光酶标仪检测GFP蛋白的结果与流式细胞术、Western-blotting的检测结果之间均呈现明显的相关性。结论：建立的利用多功能酶标仪检测GFP蛋白方法方便快捷，可以实现短时间对大样本进行检测。
목적：건립일충간단、쾌속、고통량적검측GFP단백적방법。방법：용TECAN다공능매표의Infinite M200pro검측GFP단백재불동완충액중적형광강도；용매표의검측함GFP적단백양품제도희석후적형광강도，건립형광강도여GFP상대함량지간적표준곡선；분별비교형광매표의적검측결과여류식세포술화Western-blotting검측결과적상관성。결과：사용50 mmol/L Tris-HCl(pH=10)적완충액희석GFP단백경유리우검측저농도적GFP；형광매표의검측GFP적형광독수동GFP상대함량지간적회귀계수2R>0.99。형광매표의검측GFP단백적결과여류식세포술、Western-blotting적검측결과지간균정현명현적상관성。결론：건립적이용다공능매표의검측GFP단백방법방편쾌첩，가이실현단시간대대양본진행검측。
OBJECTIVE:To establish a simple,fast and high throughput screening method for green fluorescent protein (GFP) assay.METHODS:We applied TECAN multi function measuring instrument infinite M200pro to examine the fluorescence intensity of GFP sample in different dilution buffers,and established the standard curve between fluorescence intensity and GFP relative content. Compared the results obtained from multi function measuring instrument,fluorescence-activated cell sorting (FACS) and Western-blotting.RESULTS:50 mmol/L Tris-HCl (pH=10) 2was more suitable for examining GFP samples with low content than other dilution buffers. The regression coefficientR between fluorescent value and GFP content was above 0.99 by using multi function measuring instrument. The results obtained from multi function measuring instrument correlated significantly with those obtained from FACS and Western-blotting.CONCLUSION:The established method of GFP assay using multi function measuring instrument was convenient,fast a nd could be u sed to s creen large samples in a short time.