Scalera V, Huang Y K, Hildmann B, Murer H
Membr Biochem. 1981;4(1):49-61. doi: 10.3109/09687688109065422.
Basal-lateral membranes were separated in a self-orienting Percoll (modified colloidal silica) gradient from a heavy microsomal membrane fraction by centrifugation at 48,000g for 0.5 h. The (Na+--K+)-ATPase activity as a marker enzyme for the basal-lateral plasma membrane was 20-fold enriched by this procedure. The adenylate-cyclase activity measured in the basal-lateral membrane fraction was stimulated 6-fold by parathyrin and only up to 1.5-fold by arginine-vasopressin, calcitonin, or isoproterenol. The yield of basal-lateral plasma membranes was 5 to 10 percent of the amount initially present in the homogenate. The method is also applicable to the pig kidney.
通过在48,000g下离心0.5小时，在自定向Percoll（改性胶体二氧化硅）梯度中从重微粒体膜部分分离出基底外侧膜。通过该程序，作为基底外侧质膜标记酶的（Na + -K +）-ATP酶活性富集了20倍。在基底外侧膜部分测得的腺苷酸环化酶活性，甲状旁腺素刺激其活性提高了6倍，而精氨酸加压素、降钙素或异丙肾上腺素仅使其活性提高了1.5倍。基底外侧质膜的产量为匀浆中最初存在量的5%至10%。该方法也适用于猪肾。
