以其在乳腺癌和卵巢癌中的肿瘤抑制活性而闻名,乳腺癌易感基因1(Brca1)参与包括DNA修复在内的多种细胞通路,抗氧化剂信号,凋亡,和细胞周期调控。BRCA1可以在细胞质和细胞核之间移位以发挥其各种作用。本文是用于测量全细胞裂解物(WCL)中BRCA1蛋白水平的程序,以及不同胎龄小鼠组织的核(N)和细胞质(C)部分。该方法采用多个加载对照以确保级分和总蛋白染色的适当分离,用于不同样品的更一致的比较。该方法对于识别BRCA1在各种研究领域中的缺陷和定位很有用。包括发展,神经变性,和癌症。
Known for its tumor suppressor activity in breast and ovarian cancers, the breast cancer 1 susceptibility gene (Brca1) is involved in a variety of cellular pathways including DNA repair, antioxidant signaling, apoptosis, and cell cycle regulation. BRCA1 can translocate between the cytoplasm and nucleus to perform its various roles. Herein is a procedure for measuring BRCA1 protein levels in the whole cell lysate (WCL), as well as in the nuclear (N) and cytoplasmic (C) fractions of mouse tissues at different gestational ages. The method employs multiple loading controls to ensure proper separation of fractions and a total protein stain for more consistent comparisons of dissimilar samples. This method is useful for identifying BRCA1 deficiencies and localization in a variety of research fields, including development, neurodegeneration, and cancer.