Alamar Blue-B13细胞株生物法测定IL-5水平

目的 探讨应用Alamar Blue测定细胞因子IL-5的方法。方法 将依赖IL-5的B13细胞加到细胞培养板上,加入系列稀释的待测样品IL-5/CHO细胞培养上清,孵育4d后加入Alamar Blue,继续孵育6h测定每孔A值(A_(570)-A_(600))。结果 随着样品中IL-5浓度的升高,B13细胞增殖程度增加,A值增大。敏感性较生物素ELISA法提高12～15倍。结论 AlamarBlue-B13细胞株生物法是一种测定IL-5的可行的、无放射性污染的、敏感的方法。

Alamar Blue-B13 cell line bioassay for measuring the level of interleukin-5

Objective To explore a method for detecting the level of IL-5 useing Alamar Blue. Methods B13 cells were added into the cell culture plate,followed by the addition of a series of diluted samples - the culture supernatants of IL-5/CHO cell. After incubated for 4 days, Alamar Blue was added,followed by incubation for 4 hours. Then the colorimetric changes of Alamar Blue were measured, at 570 nm and 600 nm. Results With increase of the concentration of IL-5 in the supernatants, the growth of B13 cells became beffer and the value absor-bance of higher. Conclusion The Alamar Blue-B13 cell line bioassay is a valid, non-radioactive and sensitive method for measurement of the level of IL-5.