~3H-亮氨酸掺入法测定肺泡巨噬细胞蛋白合成能力的实验方法的探讨

STUDY ON THE METHOD OF DETERMINATION OF PROTEIN SYNTHESIS BY PAM WITH 3~H-LEUCINE INCORPORATION

用测量掺入蛋白质的放射性亮氨酸的方法,研究了体外PAM蛋白合成的实验条件。结果说明加入同位素后培养2—3小时,放射性脉冲数最高。实验所用细胞密度以10~0/ml较为适宜,在此条件下所加同位素以2μci较好。此外,对其他培养条件也进行了研究。 此法简单,稳定,重现性好。

Study on PAM Protein synthesis in vitro by incorporation of 3H-leucine in protein was carried out. It demonstrated that incorporation reached to the peak cpm value 2 to 3 hours after incubation with 3H-leucine. The cell concentration. was 106/ml. Satisfactory results were obtained when 2μci/ml 3H-leu-cine was used in incorporation. Experiments at different conditions for cell culture were carried out too.This method has advantage of Simplicity and good reproducibility.