This second edition volume expands on the previous edition with a discussion of the latest methods used to explore collagen biology from molecular synthesis and folding to biomechanical analysis and tissue engineering applications. The chapters in this book are organized into four parts: Part One introduces strategies for manipulating collagen at the molecular level, including gene targeting, mutagenesis, and functional assays. Part Two focuses on cellular and biochemical aspects of collagen biology, featuring protocols to study the effects of hypoxia, assess triple helix stability, and investigate ECM-rich environments using proteomics and mass spectrometry. Part Three covers structural and imaging techniques such as atomic force microscopy and advanced visualization of collagen suprastructures, and Part Four looks at translational and biomedical research, providing protocols for constructing 3D ECM scaffolds, engineering collagen-based biomaterials, and modeling tumor microenvironments. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and comprehensive, Collagen: Methods and Protocols, Second Edition is a valuable resource for researchers in cell biology, biochemistry, tissue engineering, and translational medicine, and will support the development of novel strategies to explore and manipulate the power of collagen.