Betti Michele, Ciacci Caterina, Abramovich Sigal, Frontalini Fabrizio
Department of Biomolecular Science, Urbino University, 61029 Urbino, Italy.
Department of Earth and Environmental Sciences, Ben Gurion University of the Negev, Beer Sheva 84105, Israel.
Life (Basel). 2021 May 5;11(5):418. doi: 10.3390/life11050418.
Proteins are essential to life, and the evaluation of their content, identification, and modification represents a fundamental assay in biochemistry research. Different analytical techniques and protocols have been specifically designed but have rarely been compared. Here, we test and compare a variety of methodologies and treatments for the quantification of proteins in , a larger symbiont-bearing benthic foraminiferal species. These analyses specifically include (a) lysis buffer (homemade vs. RIPA), (b) protein assays (Lowry, BCA, and Bradford), (c) ultrasonic bath treatment, and (d) protein staining (silver staining vs. Coomassie blue). On the basis of the comparative outcome, we suggest using the homemade lysis buffer, Lowry or BCA assays, ultrasonic bath treatment, and silver stain to maximize the extraction and characterization of protein for . This protocol might be suitable and extended to other benthic foraminiferal species, including the smaller ones.
蛋白质对生命至关重要，对其含量、鉴定和修饰的评估是生物化学研究中的一项基本测定。已经专门设计了不同的分析技术和方案，但很少进行比较。在这里，我们测试并比较了多种方法和处理方式，用于对一种较大的含共生体底栖有孔虫物种中的蛋白质进行定量分析。这些分析具体包括：(a) 裂解缓冲液（自制与RIPA），(b) 蛋白质测定法（洛瑞法、BCA法和布拉德福德法），(c) 超声浴处理，以及(d) 蛋白质染色（银染与考马斯亮蓝染色）。基于比较结果，我们建议使用自制裂解缓冲液、洛瑞法或BCA法测定、超声浴处理和银染，以最大限度地提取和鉴定该物种的蛋白质。该方案可能适用于并扩展到其他底栖有孔虫物种，包括较小的物种。
