Park Jun, Mabuchi Masaharu, Sharma Ajay
Millipore Corporation, 17 Cherry Hill Drive, Danvers, MA, 01923, USA.
Methods Mol Biol. 2009;536:527-31. doi: 10.1007/978-1-59745-542-8_53.
In 1988, two separate investigators reported a novel method of detecting unstained protein bands on polyvinylidene fluoride (PVDF) membranes using white light transillumination. This simple method exploits the intrinsic hydrophobicity of PVDF membrane, which enables the visual observation of transferred protein bands due to differential wetting patterns between protein bands and the membrane itself. This method applies only to hydrophobic PVDF membranes, because hydrophilic membranes such as nitrocellulose wet out completely, rendering the protein bands invisible by transillumination. Transilluminational protein visualization can detect submicrogram quantities of proteins while circumventing the use of protein stains, which can potentially interfere with downstream analysis such as N-terminal sequencing. In this chapter, we demonstrate efficient transilluminational protein visualization on a recently introduced low-fluorescence PVDF membrane, normally used for downstream fluorescent immunodetection.
1988年，两位独立的研究者报告了一种利用白光透照检测聚偏二氟乙烯（PVDF）膜上未染色蛋白条带的新方法。这种简单的方法利用了PVDF膜固有的疏水性，由于蛋白条带与膜本身之间的润湿性差异，使得转移的蛋白条带能够被肉眼观察到。该方法仅适用于疏水的PVDF膜，因为诸如硝酸纤维素等亲水膜会完全湿透，通过透照使蛋白条带不可见。透照蛋白可视化能够检测到亚微克量的蛋白质，同时避免使用可能干扰诸如N端测序等下游分析的蛋白质染色剂。在本章中，我们展示了在一种最近推出的低荧光PVDF膜上进行高效的透照蛋白可视化，这种膜通常用于下游荧光免疫检测。
