鞘氨醇-1-磷酸的定量测定——一种新的竞争性结合方法

Quantitative Measurement of Sphingosine-1-phosphate : A Novel Competitive Binding Method

鞘氨醇 1 磷酸 (SPP)是重要的细胞第二信使 ,影响细胞的生长和死亡 .通过培养和收集转染SPP受体 EDG 1的HEK2 93细胞 ,与标记及非标记SPP共孵育 ,利用它们与HEK2 93细胞的竞争性结合 ,测定细胞、血清和组织中SPP含量 .该法无需特殊仪器 ,可以测到皮摩尔水平的低含量 ,批间差异小于 15 % (6次 ) .

Sphingosine 1 phosphate(SPP) is an important second messenger involved in cell growth and cell death. HEK293 cells transcripted with EDG 1, a SPP receptor, were cultured and harvested, and incubated with 32 P labeled or non labeled SPP. The SPP level was determined based on competitive binding of SPP or 32 P SPP to HEK cells. This method does not require special apparatus, and able to measure the content of SPP as low as pmol with high sensitivity and good repetition. The between groups deviation was less than 15%.