Palecek J, Ubbels G A, Mácha J
Histochemistry. 1982;76(4):527-38. doi: 10.1007/BF00489907.
Tubulin can be isolated and purified from Xenopus laevis eggs through modification of Olmstedt's (1970) tubulin isolation method, viz. by repeating the vinblastin precipitation step after resuspension of the sediment in a detergent-containing stabilizing medium. By this we overcome the deleterious influence of the yolk granules in the isolation procedure. From 11 of Xenopus laevis eggs 25 mg VB-paracrystals can be obtained. The apparent molecular weight of the purified tubulin is 52,800. Antiserum against the purified Xenopus VB-paracrystals, raised in 2 Chinchilla rabbits, cross-reacts in immunodiffusion tests in agar gels with rat brain tubulin and with tubulin isolated from Xenopus laevis eggs by the described procedure. Specific indirect fluorescence staining and appropriate control reactions reveal that cilia of Tetrahymena pyriformis, cytoplasmic networks in cultured mouse Leydig cells, as well as mitotic spindles and nuclear regions in paraffin sections of Xenopus laevis blastulae, react with the antibodies against Xenopus laevis egg tubulin as well as with monoclonal antibodies against pig brain tubulin. These results provide additional evidence for the view that tubulin antibodies are neither species nor tissue specific and show that under appropriate conditions tubulin containing structures can be visualized in paraffin sections.
微管蛋白可通过对奥姆施泰特（1970年）的微管蛋白分离方法进行改进，从非洲爪蟾卵中分离和纯化，即：在将沉淀物重悬于含去污剂的稳定介质后，重复长春花碱沉淀步骤。通过这种方法，我们克服了卵黄颗粒在分离过程中的有害影响。从11个非洲爪蟾卵中可获得25毫克长春花碱副晶体。纯化后的微管蛋白的表观分子量为52,800。在2只青紫蓝兔中制备的针对纯化的非洲爪蟾长春花碱副晶体的抗血清，在琼脂凝胶免疫扩散试验中与大鼠脑微管蛋白以及通过所述方法从非洲爪蟾卵中分离的微管蛋白发生交叉反应。特异性间接荧光染色和适当的对照反应表明，梨形四膜虫的纤毛、培养的小鼠睾丸间质细胞中的细胞质网络以及非洲爪蟾囊胚石蜡切片中的有丝分裂纺锤体和核区域，与针对非洲爪蟾卵微管蛋白的抗体以及针对猪脑微管蛋白的单克隆抗体发生反应。这些结果为微管蛋白抗体既非物种特异性也非组织特异性的观点提供了额外证据，并表明在适当条件下，含微管蛋白的结构可在石蜡切片中显现出来。
