For proteome analyses, the tissue samples are mostly preserved either snap frozen or formalin-fixed, paraffin-embedded form. Use of RNAlater—a non-toxic solution primarily used to stabilize the RNA content of samples—in tissue preservation for proteome analysis recently described equally reliable with snap-frozen preservation in human tissues. Even though RNALater storage has great potential in the preservation of Peripheral Blood Mononuclear Cells (PBMC), its impact on the results of proteome analysis is poorly described at qualitative and quantitative measures. The present study investigated protein profiles of RNAlater preserved and fresh PBMCs using three extraction buffers viz. Triton X-100, RIPA and SDS. Proteins are separated in SDS-PAGE and quantified using densitometry. On an average 19.3 bands from fresh and 15.6 bands from RNAlater storage cells were obtained with a molecular weight ranging from 25 to > 250 kDa. RNAlater storage generated a fewer number and lesser quantity of low molecular weight proteins while yielded a similar or high quantity of high molecular weight protein fractions. The principal component analysis showed that Triton X-100 is inferior as compared to SDS and RIPA with respect to their protein bands and quantity yielded. While RNAlater is effective in preserving PBMC for proteome analysis, our findings warrant caution in its use in proteomics experiments especially if the target is low molecular weight proteins.

中文翻译：

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使用不同的蛋白质提取缓冲液评估RNAlater存储和新鲜PBMC细胞的蛋白质谱。

对于蛋白质组分析，组织样品大多保存为速冻或福尔马林固定，石蜡包埋形式。RNAlater（一种主要用于稳定样品RNA含量的无毒溶液）在组织保存中用于蛋白质组分析的方法最近被描述为与人体组织速冻保存同样可靠。尽管RNALater储存在外周血单个核细胞（PBMC）的保存方面具有巨大潜力，但在定性和定量措施上，其对蛋白质组分析结果的影响却很少。本研究使用三种提取缓冲液来研究RNAlater保存和新鲜PBMC的蛋白质谱。Triton X-100，RIPA和SDS。在SDS-PAGE中分离蛋白质，并使用光密度法定量。平均而言，新鲜的和新鲜的有15.3条。从RNAlater存储细胞获得了6条带，分子量为25至> 250 kDa。RNAlater储存产生较少数量和较少数量的低分子量蛋白质，而产生相似或大量的高分子量蛋白质馏分。主成分分析表明，就蛋白质条带和产量而言，Triton X-100比SDS和RIPA差。尽管RNAlater可以有效地保留PBMC用于蛋白质组分析，但我们的发现值得在蛋白质组学实验中谨慎使用，尤其是当目标是低分子量蛋白质时。RNAlater储存产生较少数量和较少数量的低分子量蛋白质，而产生相似或大量的高分子量蛋白质馏分。主成分分析表明，就蛋白质条带和产量而言，Triton X-100比SDS和RIPA差。尽管RNAlater可以有效地保留PBMC用于蛋白质组分析，但我们的发现值得在蛋白质组学实验中谨慎使用，尤其是当目标是低分子量蛋白质时。RNAlater储存产生较少数量和较少数量的低分子量蛋白质，而产生相似或大量的高分子量蛋白质馏分。主成分分析表明，就蛋白质条带和产量而言，Triton X-100比SDS和RIPA差。尽管RNAlater可以有效地保留PBMC用于蛋白质组分析，但我们的发现值得在蛋白质组学实验中谨慎使用，尤其是当目标是低分子量蛋白质时。