Murray Christopher I, Barrett Meagan, Van Eyk Jennifer E
Department of Physiology, Queen's University, Kingston, Canada.
Proteomics. 2009 Aug;9(15):3934-8. doi: 10.1002/pmic.200701170.
The nuclear fraction of the ProteoExtract subcellular fractionation kit was assessed using frozen rat liver and heart tissue. Fractionation was evaluated by Western blot using protein markers for various subcellular compartments and followed up with LC/MS/MS analysis of the nuclear fractions. Of the proteins identified, nuclear proteins were in the minority (less than 15%) and there was poor representation of the various nuclear substructures when compared with liver nuclear isolations using a classical density-based centrifugation protocol. The ProteoExtract kit demonstrated poor specificity for the nucleus and offers limited promise for proteomics investigations of the nuclear subproteome in frozen tissue samples.
使用冷冻大鼠肝脏和心脏组织对ProteoExtract亚细胞分级分离试剂盒的细胞核组分进行评估。通过蛋白质印迹法，使用针对各种亚细胞区室的蛋白质标志物对分级分离进行评估，并对细胞核组分进行液相色谱串联质谱（LC/MS/MS）分析。在鉴定出的蛋白质中，核蛋白占少数（不到15%），与使用经典密度梯度离心方案进行的肝脏细胞核分离相比，各种核亚结构的代表性较差。ProteoExtract试剂盒对细胞核的特异性较差，在冷冻组织样本中对核亚蛋白质组进行蛋白质组学研究的前景有限。
