Ninomiya M, Suganuma M, Paik N S, Muto Y, Fujiki H
National Cancer Center Research Institute, Tokyo, Japan.
FEBS Lett. 1988 Jun 20;233(2):255-8. doi: 10.1016/0014-5793(88)80437-x.
The specific binding of [3H]retinoids to cellular retinoid-binding proteins was measured directly by the cold acetone filtration method. After incubation of purified cellular retinoid-binding proteins with [3H]retinoids with or without competitors for 2-4 h, bound ligands were separated from free by filtration using cold acetone. Nonspecific binding of the ligands was reduced sufficiently to allow measurement of specific binding of [3H]retinoids to cellular retinoid-binding proteins. This method has the advantages of being rapid and practical and giving reproducible results.
通过冷丙酮过滤法直接测定[3H]类视黄醇与细胞类视黄醇结合蛋白的特异性结合。将纯化的细胞类视黄醇结合蛋白与[3H]类视黄醇在有或无竞争者的情况下孵育2 - 4小时后，通过冷丙酮过滤将结合的配体与游离的配体分离。配体的非特异性结合被充分降低，从而能够测定[3H]类视黄醇与细胞类视黄醇结合蛋白的特异性结合。该方法具有快速、实用且结果可重复的优点。
