The Nuclear Membrane

This book compiles protocols that encompass a diverse range of classical and contemporary approaches, including biochemical techniques, proteomic analyses, microscopic observations, and biophysical measurements. Written in the highly successful Methods in Molecular Biology series format, the chapters include brief introductions to the material, lists of necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section which highlights **tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, The Nuclear Membrane: Methods and Protocols aims to be a valuable resource for scientists seeking to explore the nuclear structure and functions in the postgenomic era.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Lipidomic profiling of isolated nuclei from mice and cultured cells.- Profiling plant nuclear envelope composition using subtractive proteomics.- Ultrastructural visualization of the nuclearenvelope in HeLa cells.- Imaging nuclear envelopes using correlative AFM/fluorescence microscopy.- Fast super-resolution live cell imaging of nuclear membrane-endoplasmic reticulum dynamics Immunostaining the yeast nuclear membrane for imaging by super-resolution fluorescence microscopy.- Visualizing phosphatidic acid and diacylglycerol at the nuclear envelope in fission yeast.- Measuring molecular mass densities at subcellular resolution using optical diffraction tomography.- Quantification and comparison of protein distribution on nuclear membrane.- Quantifying nuclear shape fluctuations along the cell cycle, with a focus on early mitosis.- Micropost arrays to model ECM fiber obstacles during cell migration in confinement.- Use of nucleoporin-conjugated beads to study the nuclear pore complex assembly on the nuclear membrane.- T4 DNA-induced reconstruction of artificial nuclei in living mouse oocyte.- ** Preparation of nucleoplasmic extract and its application in DNA end processing.-Liposome floatation assays to study membrane interactions of nucleoporins.- ** Giant unilamellar vesicles (GUVs) to study membrane interaction of nucleoporins.- One-pot reconstitution of GPCRs into unilamellar vesicles for fluorescence-based phospholipid scramblase activity assay.- Identifying genomic DNA sequences near the nuclear lamina using proximity biotinylation with Ascorbate Peroxidase.- ** Pairing LaminB1-DamID and Immuno-3D-FISH to resolve and verify peripheral genome organization of adipogenesis.-Measurement of spatial contact map using sequential FISH.**