histone h2b anti rabbit antibody  (Proteintech)

Journal: Nature Communications

Article Title: Revealing the biophysics of lamina-associated domain formation by integrating theoretical modeling and high-resolution imaging

doi: 10.1038/s41467-025-63244-1

Figure Lengend Snippet: A Representative Voronoi density rendering of STORM images of hMSC nuclei, color-coded to indicate histone H2B density levels (blue for low density and red for high density). B The distribution of heterochromatin domain sizes obtained from STORM images in interior (Top) and at periphery (Bottom). Notably the distributions show a characteristic mean size. C Chromatin-chromatin interactions result in segregation of chromatin into euchromatin and heterochromatin phases. These interactions incorporate chromatin-chromatin interactions, including those mediated by crosslinking molecules such as HP1α, as well as the direct interactions between segments of chromatin. The model also includes the interactions between chromatin and the lamina, mediated by anchoring proteins such as LAP2α/β and LBR. These chromatin-lamina interactions, localized at the nuclear periphery, result in the formation of heterochromatin rich lamina-associated domains (LADs). D Epigenetic factors, such as HDAC and HMT, regulate acetylation and methylation reactions that allow interconversion of heterochromatin and euchromatin phases, captured via first-order reaction kinetics. The diffusion of water and epigenetic markers is included in the continuum model. The anchoring of chromatin to the nuclear lamina is also mediated by HDAC3 , . E The contour plot of free energy density shows the two wells (local minima) corresponding to the two stable phases of chromatin – euchromatin (blue) and heterochromatin (red). We schematically show how an initial homogeneous distribution of chromatin (white circle) will evolve towards the two energy wells.

Article Snippet: The cells were then plated in basal growth medium, which consisted of high-glucose DMEM medium supplemented with 10% penicillin–streptomycin, L-glutamine, and 10% FBS. hMSCs and hTCs were fixed using methanol-ethanol (1:1) at −20 °C for 6 min, followed by blocking with a solution of 10%(w/v) bovine serum albumin (BSA) in phosphate-buffered saline (PBS) for 1 h . Subsequently, the cells were subjected to overnight incubation at 4 °C with a 1:50 dilution of histone H2B anti-rabbit antibody (ProteinTech, #15857-1-AP).

Techniques: Methylation, Diffusion-based Assay

Journal: Nature Communications

Article Title: Revealing the biophysics of lamina-associated domain formation by integrating theoretical modeling and high-resolution imaging

doi: 10.1038/s41467-025-63244-1

Figure Lengend Snippet: A Representative Voronoi density rendering of H2B STORM images, color-coded to indicate density levels (blue for low density and red for high density). B Steady-state chromatin organization predicted by numerical simulations showing phase separation into compacted heterochromatin domains (red) and loosely packed euchromatin domains (blue) and the formation of heterochromatin-rich LADs at the nuclear periphery. Numerical simulations show the synergistic effects of chromatin-lamina affinity and methylation rates on LADs and inner heterochromatin domain morphology. At low levels of chromatin-lamina affinity, as the methylation level increases, the radius of the discrete LADs increases at a fixed value of the contact angle \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$(\theta )$$\end{document} ( θ ) . The morphology of LADs is determined by both the level of methylation and chromatin-lamina affinity. Low affinity results in the formation of isolated LADs, while at greater levels of affinity LAD spread along the lamina. C Schematic illustrating the morphology of LAD at small level of chromatin-lamina affinity. D The scaling relationship between the thickness of LAD and chromatin-lamina affinity at a given methylation level showing three distinct regimes. E The scaling relation between the thickness of LAD and chromatin-lamina affinity at varying methylation levels. F Workflow of the theoretical framework for extracting methylation rates and chromatin-lamina affinity by integrating super resolution STORM images of DNA with theoretical analyses. Within the interior of the nucleus, STORM image analysis gives the distribution of heterochromatin domain radii ( R d ), and at periphery, it provides the distribution of LAD thickness \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${{T}}_{{LAD}}$$\end{document} T L A D . The combination of these distributions with the theoretical framework predicts the corresponding distribution of histone methylation rates \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\Gamma }_{{me}}$$\end{document} Γ m e and the chromatin-lamina affinities \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${V}_{{LAD}}$$\end{document} V L A D . Notably the distribution of \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${V}_{{LAD}}$$\end{document} V L A D is bimodal. The red curve in all plots shows the smoothed density plot. All source data are provided as a Source Data file.

Article Snippet: The cells were then plated in basal growth medium, which consisted of high-glucose DMEM medium supplemented with 10% penicillin–streptomycin, L-glutamine, and 10% FBS. hMSCs and hTCs were fixed using methanol-ethanol (1:1) at −20 °C for 6 min, followed by blocking with a solution of 10%(w/v) bovine serum albumin (BSA) in phosphate-buffered saline (PBS) for 1 h . Subsequently, the cells were subjected to overnight incubation at 4 °C with a 1:50 dilution of histone H2B anti-rabbit antibody (ProteinTech, #15857-1-AP).

Techniques: Methylation, Isolation