Raykin Julia, Snider Eric, Bheri Sruti, Mulvihill John, Ethier C Ross
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, United States.
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, United States; Department of Mechanical and Manufacturing Engineering, School of Engineering, Trinity College Dublin, Dublin, Ireland.
Anal Biochem. 2017 Mar 15;521:8-10. doi: 10.1016/j.ab.2017.01.003. Epub 2017 Jan 6.
Gelatinase zymography is a commonly used laboratory procedure; however, variability in sample loading and concentration reduce the accuracy of quantitative results obtained from this technique. To facilitate normalization of gelatinase activity by loaded protein amount, we developed a protocol using the trihalocompound 2,2,2-trichloroethanol to allow for gelatin zymography and total protein labeling within the same gel. We showed that detected protein levels increased linearly with loading, and describe a loading concentration range over which normalized gelatinase activity was constant. We conclude that in-gel total protein detection is feasible in gelatin zymography and greatly improves comparison of gelatinase activity between samples.
明胶酶谱法是一种常用的实验室方法；然而，样品上样量和浓度的变异性会降低通过该技术获得的定量结果的准确性。为便于根据上样蛋白量对明胶酶活性进行标准化，我们开发了一种方案，使用三卤化合物2,2,2-三氯乙醇，以便在同一凝胶内进行明胶酶谱分析和总蛋白标记。我们发现检测到的蛋白水平随上样量呈线性增加，并描述了标准化明胶酶活性保持恒定的上样浓度范围。我们得出结论，在明胶酶谱分析中进行凝胶内总蛋白检测是可行的，并且极大地改善了样品间明胶酶活性的比较。
