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Titlebook: Cell Cycle Oscillators; Methods and Protocol Amanda S. Coutts,Louise Weston Book 2021Latest edition Springer Science+Business Media, LLC, p

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Phosphatase and Kinase Substrate Specificity Profiling with Pooled Synthetic Peptides and Mass Spec. Determining the inherent substrate specificity of kinases and phosphatases is essential for understanding their cellular roles. Synthetic peptides have long served as substrate proxies for defining intrinsic kinase and phosphatase specificities. Here, we describe a high throughput protocol to simu
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Whole-Mount Immunostaining for the Identification of Histone Modifications in the S-Phase Nuclei ofevious studies have demonstrated that dramatic changes in local chromatin structure are required for the initiation and progression of DNA replication, and that histone modifications play an essential role in the determination of chromatin structure in S phase. Since euchromatic and heterochromatic
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Optimizing Cell Synchronization Using Nocodazole or Double Thymidine Block,eve the cell culture synchronization but not all type of cells respond equally to a given concentration of these drugs. Here we describe a simple optimization method to select concentrations and timings for nocodazole or thymidine treatments using fluorescence staining. In addition, we provide detai
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Elucidating Human Mitosis Using an Anaphase-Like Cell-Free System,ory and structural proteins. These series of events ultimately secure genome stability and time-invariant cellular properties during cell proliferation. Two of the core enzymes regulating mitotic milestones in all eukaryotes are cyclin dependent kinase 1 (CDK1) with its coactivator cyclin B, and the
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,EDU (5-Ethynyl-2′-Deoxyuridine)-Coupled Fluorescence-Intensity Analysis: Determining Absolute Paramycle stages (G1, S, and G2) are detailed herein. This methodology utilizes flow cytometry to take full advantage of the excellent stoichiometric properties of click chemistry. This allows detection, via azide-fluorochrome coupling, of the modified deoxynucleoside 5-ethynyl-2′-deoxyuridine (EDU) inco
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