Titlebook: Cell Polarity Signaling; Methods and Protocol Chenbei Chang,Jianbo Wang Book 2022 Springer Science+Business Media, LLC, part of Springer Na

BET

Malfunction

impaction

Use of Fluorescence Recovery After Photobleaching (FRAP) to Measure In Vivo Dynamics of Cell Junctiit relatively slow junctional turnover. We provide a step-by-step practical guide to performing FRAP, and list a series of controls and optimizations to do before conducting a FRAP experiment. Finally, we describe how to present the FRAP data for publication.

Prologue

FRET Imaging of Rho GTPase Activity with Red Fluorescent Protein-Based FRET Pairs,iotemporal resolution in living cells and organisms. FRET can be detected in appropriately configured microscopes as changes in fluorescence intensity, lifetime, and anisotropy. Here, we describe the preparation of samples expressing FP-based FRET sensors for RhoA kinase, intensity- and lifetime-based FRET imaging, and postimaging data analysis.

Metastasis

Single-Cell Single-Molecule Pull-Down (sc-SiMPull) for Detection of Protein Complexes from Embryonigle-molecule pull-down (sc-SiMPull), a TIRF microscopy-based coimmunoprecipitation method, to visualize thousands of individual proteins, their binding partners, and protein complex stoichiometry directly from single-cell lysate. By iterating sc-SiMPull over time, temporal dynamics of protein complexes in response to signaling can be constructed.

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Characterization of Axon Guidance Phenotypes in Wnt/PCP Mutant Mice,d a variety of axon tracing and labeling techniques, including immunohistochemistry (IHC), DiI, BDA, and fluorescent reporters (GFP or tdTomato). These studies have primarily been conducted in spinal cord commissural axons, but have been applied to retinal ganglion cell axons, corticospinal tract axons, dopaminergic and serotonergic projections.

Middle-Ear

Chenbei Chang,Jianbo WangIncludes cutting-edge techniques.Provides step-by-step detail essential for reproducible results.Contains key implementation advice from the experts

平息

Methods in Molecular Biologyhttp://image.papertrans.cn/c/image/222862.jpg