Hatch G E, Nichols W K, Hill H R
J Immunol Methods. 1978;19(4):301-8. doi: 10.1016/0022-1759(78)90014-5.
Rapid treatment of leukocyte suspensions was found to be an effective alternative to acid treatment in the preparation of these cells for radioimmunoassay of cyclic GMP and cyclic AMP. A 2-sec heating to boiling temperature, followed by sonication and micropore filtration, was employed. This procedure adequately inactivated or removed enzymes and binding proteins that can alter cyclic nucleotide concentrations or otherwise interfere with the radioimmunoassay. Moreover, heating in this manner did not appear to affect the stability of cyclic nucleotides or cause significant formation of cyclic GMP from endogenous GTP. Recovery of cyclic nucleotides after heating and filtration was high (89-96%), making possible the measurement of both cyclic GMP and cyclic AMP in small cellular samples. Variation in cyclic nucleotide recovery was small (+/- 2-4% SD), therefore individual recovery determinations were unnecessary.
在制备用于环磷酸鸟苷（cGMP）和环磷酸腺苷（cAMP）放射免疫测定的细胞时，发现对白细胞悬液进行快速处理是酸处理的一种有效替代方法。采用将细胞悬液加热至沸腾温度2秒，随后进行超声处理和微孔过滤。该程序能充分灭活或去除可改变环核苷酸浓度或以其他方式干扰放射免疫测定的酶和结合蛋白。此外，以这种方式加热似乎不会影响环核苷酸的稳定性，也不会导致内源性鸟苷三磷酸（GTP）大量形成环磷酸鸟苷。加热和过滤后环核苷酸的回收率很高（89 - 96%），使得在小细胞样本中同时测量环磷酸鸟苷和环磷酸腺苷成为可能。环核苷酸回收率的变化很小（±2 - 4%标准差），因此无需进行个体回收率测定。
