Collagen type I is the most abundant extracellular matrix protein, and collagen type I supramolecular assemblies (e.g., tissue grafts, biomaterials and cell-assembled systems) are used extensively in tissue engineering and regenerative medicine. Many studies, for convenience or economic reasons, do not accurately determine collagen type I purity, concentration, solubility and extent of cross-linking in biological specimens, frequently resulting in erroneous conclusions. In this protocol, we describe solubility; normal, reduced and delayed (interrupted) SDS-PAGE; hydroxyproline; Sircol collagen and Pierce BCA protein; denaturation temperature; ninhydrin/trinitrobenzene sulfonic acid; and collagenase assays and assess them in a diverse range of biological samples (e.g., tissue samples; purified solutions or lyophilized materials; 3D scaffolds, such as sponges and hydrogels; and cell media and layers). Collectively, the described protocols provide a comprehensive, yet fast and readily implemented, toolbox for collagen type I characterization in any biological specimen.

中文翻译：

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实验工具箱，用于表征生物样本中的I型哺乳动物胶原蛋白。

I型胶原蛋白是最丰富的细胞外基质蛋白，I型胶原蛋白的超分子组装体（例如组织移植物，生物材料和细胞组装系统）广泛用于组织工程和再生医学中。出于方便或经济原因，许多研究未能准确确定生物样品中I型胶原蛋白的纯度，浓度，溶解度和交联程度，经常会得出错误的结论。在此协议中，我们描述了溶解度；正常，减少和延迟（中断）的SDS-PAGE；羟脯氨酸 Sircol胶原蛋白和Pierce BCA蛋白；变性温度 茚三酮/三硝基苯磺酸; 和胶原酶分析，并在各种生物样品（例如组织样品；纯化的溶液或冻干的材料； 3D支架，如海绵和水凝胶；以及细胞培养基和层）。总体而言，所描述的方案为任何生物学标本中的I型胶原蛋白表征提供了一个全面而又快速且易于实施的工具箱。