Contribution of cell culture additives to the two-dimensional protein patterns of mouse macrophages
Contribution of cell culture additives to the two-dimensional protein patterns of mouse macrophages
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DOI：
10.1002/elps.200500744
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发表时间：
2006-04-01
期刊：
影响因子：
2.9
通讯作者：
Gemeiner, Manfred
中科院分区：
文献类型：
--
作者：
Miller, Ingrid;Radwan, Marta;Gemeiner, Manfred
在原代小鼠巨噬细胞（骨髓源性巨噬细胞（BMM）和腹腔巨噬细胞（PM））制备物中，追踪了用作细胞培养基中蛋白质补充剂的低水平胎牛血清（FCS）。利用差异图像凝胶电泳和免疫印迹技术对该添加剂的主要成分进行了双向电泳图谱分析。细胞制备中的额外洗涤步骤有助于将四种最高丰度的胎血清蛋白（血清白蛋白（SA）、α（1）-甲胎蛋白（AFP）、α（1）-抗胰蛋白酶（α（1）AT）和转铁蛋白（Tf））的水平降低至总蛋白的1%以下。平行记录巨噬细胞斑点模式，并显示出很小的变化。呈现的结果应该是具有类似背景的细胞制剂的普遍兴趣。
Low levels of fetal calf serum (FCS), used as protein supplement in cell culture medium, were traced in preparations of primary murine macrophages (bone-marrow-derived macrophages (BMM) and peritoneal macrophages (PM)). Main components of this common additive were mapped in 2-DE by means of differential image gel electrophoresis and immunoblotting. Additional washing steps in cell preparation helped to decrease the levels of the four highest abundance foetal serum proteins (serum albumin (SA), alpha(1)-fetoprotein (AFP), alpha(1)-antitrypsin (alpha(1)AT) and transferrin (Tf)) to less than 1% of total protein. Macrophage spot pattern was recorded in parallel and showed little variation. Results presented are supposed to be of general interest for cell preparations with similar background.