Dewhurst S, Stevenson M, McComb R D, Volsky D J
Acta Neuropathol. 1987;73(4):383-6. doi: 10.1007/BF00688264.
The expression of glial fibrillary acidic protein (GFAP) in continuous glioma and non-glioma cell lines was evaluated by molecular hybridization and immunocytochemistry. RNA transcripts from the GFAP gene were detected in two of six cell lines by Northern blot analysis of cellular RNAs using a cloned mouse GFAP cDNA probe. The probe recognized a single GFAP RNA species of 2.7 kilobases under high-stringency washing conditions. The presence of GFAP transcripts as determined by molecular hybridization corresponded to the immunocytochemical detection of GFAP using two different monoclonal antibodies to this protein. These data indicate that GFAP expression in human cells can be detected by molecular hybridization using a murine GFAP cDNA probe, illustrating the high degree of inter-species conservation of GFAP. Molecular hybridization represents a reliable and sensitive method for the detection of GFAP expression in vitro, which can be used in conjunction with or as an alternative to immunocytochemistry.
通过分子杂交和免疫细胞化学方法评估了胶质纤维酸性蛋白(GFAP)在连续性胶质瘤细胞系和非胶质瘤细胞系中的表达。使用克隆的小鼠GFAP cDNA探针,通过对细胞RNA进行Northern印迹分析,在六个细胞系中的两个中检测到了GFAP基因的RNA转录本。在高严格洗涤条件下,该探针识别出一种2.7千碱基的单一GFAP RNA种类。通过分子杂交确定的GFAP转录本的存在与使用针对该蛋白的两种不同单克隆抗体进行的GFAP免疫细胞化学检测结果一致。这些数据表明,使用鼠源GFAP cDNA探针通过分子杂交可检测人细胞中的GFAP表达,这说明了GFAP在种间具有高度的保守性。分子杂交是一种用于体外检测GFAP表达的可靠且灵敏的方法,可与免疫细胞化学联合使用或作为其替代方法。