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[求助]
求教！做western blot实验的抗体如何选择？
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我想做药物作用于细胞后，细胞信号传导途径的Wnt途径中的靶基因cyclin D1的表达情况，cyclin D1是作用于G1期的重要的细胞周期蛋白，但是在抗体选择方面， 不知道cyclin D1（L283）PAb 与cyclin D1（D86）这两种抗体有何不同？是否都可以用于Wnt途径中的靶基因cyclin D1的表达情况？L和D指的是什么？ 这是两种抗体的简要说明： cyclin D1（L283）PAb The proliferation of eukaryotic cells is controlled at specific points in the cell cycle, particularly at the G1 to S and the G2 to M transitions. It is well established that the Cdc2 p34-cyclin B protein kinase plays a critical role in the G2 to M transition while cyclin A associates with Cdk2 p33 and functions in S phase. Considerable effort directed towards the identification of G1 cyclins has led to the isolation of cyclin D, cyclin C and cyclin E. Of these, cyclin D corresponds to a putative human oncogene, designated PRAD1, which maps at the site of the Bcl1 rearrangement in certain lymphomas and leukemias. Two additional human type D cyclins, as well as their mouse homologs, have been identified. Evidence has established that members of the cyclin D family function to regulate phosphorylation of the retinoblastoma gene product, thereby activating E2F transcription factors. cyclin D1（D86） During each cell cycle cyclins undergo periodic accumulation and destruction. As key regulators of the cell cycle the cyclins control important transitions by acting as regulatory subunits of the Cdks. Early in the G1 phase of the cell cycle, cyclin D1 induction is followed by cyclin E induction. This sequential progression is marked early on in G1 by the activation of Cdk4 and in mid to late G1 by the activation of Cdk2 and the hyperphosphorylation of pRB. The final transition into S phase is thought to be dependent on the increased expression and association of cyclin E and Cdk2. |
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