Cancer Biology

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    Protocols in Current Issue
    Conjugation of Fab’ Fragments with Fluorescent Dyes for Single-molecule Tracking on Live Cells
    Authors:  I-Ting Teng, Xiangning Bu and Inhee Chung, date: 09/20/2019, view: 515, Q&A: 0
    [Abstract] Our understanding of the regulation and functions of cell-surface proteins has progressed rapidly with the advent of advanced optical imaging techniques. In particular, single-molecule tracking (SMT) using bright fluorophores conjugated to antibodies and wide-field microscopy methods such as total internal reflection fluorescence microscopy have ...
    GFP-Grb2 Translocation Assay Using High-content Imaging to Screen for Modulators of EGFR-signaling
    Authors:  Julia Petschnigg and Robin Ketteler, date: 09/05/2017, view: 4019, Q&A: 0
    [Abstract] High-content screening is a useful tool to understand complex cellular processes and to identify genes, proteins or small molecule compounds that modulate such pathways. High-content assays monitor the function of a protein or pathway by visualizing a change in an image-based readout, such as a change in the localization of a reporter protein. ...
    Non-radioactive LATS in vitro Kinase Assay
    Authors:  Audrey W. Hong and Kun-Liang Guan, date: 07/20/2017, view: 6524, Q&A: 1
    [Abstract] This protocol describes a method to directly measure LATS activity by an in vitro kinase assay using YAP as a substrate.
    Generation of a Cellular Reporter for Functional BRD4 Inhibition
    Authors:  Sara Sdelci and Stefan Kubicek, date: 07/05/2017, view: 3414, Q&A: 0
    [Abstract] The ubiquitously expressed bromodomain-containing protein 4 (BRD4) is an epigenetic reader, which recruits transcriptional regulatory complexes to acetylated chromatin. Because of its role in enhancing proliferation, BRD4 has become a therapeutic target in oncology, as the inhibition of this protein leads to the reduction of the growth of many ...
    Fluorometric Estimation of Glutathione in Cultured Microglial Cell Lysate
    Authors:  Vikas Singh, Ruchi Gera, Mahaveer Prasad Purohit, Satyakam Patnaik and Debabrata Ghosh, date: 06/05/2017, view: 6303, Q&A: 6
    [Abstract] Glutathione is one of the major antioxidant defense components present in cells. It is predominantly present as reduced glutathione (GSH) and converted into oxidized glutathione (GSSG) while reducing the free radicals like hydroxyl ions (OH-). For the measurement of GSH and GSSG, o-phthalaldehyde (OPT) has been used as a fluorescent ...
    Analysis of in vivo Interaction between RNA Binding Proteins and Their RNA Targets by UV Cross-linking and Immunoprecipitation (CLIP) Method
    Authors:  Pamela Bielli and Claudio Sette, date: 05/20/2017, view: 5658, Q&A: 0
    [Abstract] RNA metabolism is tightly controlled across different tissues and developmental stages, and its dysregulation is one of the molecular hallmarks of cancer. Through direct binding to specific sequence element(s), RNA binding proteins (RBPs) play a pivotal role in co- and post-transcriptional RNA regulatory events. We have recently demonstrated that, ...
    Virtual Screening of Transmembrane Serine Protease Inhibitors
    Authors:  Antti Poso, Topi Tervonen and Juha Klefström, date: 04/20/2017, view: 4740, Q&A: 0
    [Abstract] The human family of type II transmembrane serine proteases includes 17 members. The defining features of these proteases are an N-terminal transmembrane domain and a C-terminal serine protease of the chymotrypsin (S1) fold, separated from each other by a variable stem region. Recently accumulated evidence suggests a critical role for these ...
    Differential Salt Fractionation of Nuclei to Analyze Chromatin-associated Proteins from Cultured Mammalian Cells
    Authors:  Christin Herrmann, Daphne C. Avgousti and Matthew D. Weitzman, date: 03/20/2017, view: 9079, Q&A: 0
    [Abstract] Nucleosomes are the core units of cellular chromatin and are comprised of 147 base pairs (bp) of DNA wrapped around an octamer of histone proteins. Proteins such as chromatin remodelers, transcription factors, and DNA repair proteins interact dynamically with chromatin to regulate access to DNA, control gene transcription, and maintain genome ...
    RNA-protein UV-crosslinking Assay
    Authors:  Dipak Kumar Poria and Partho Sarothi Ray, date: 03/20/2017, view: 8976, Q&A: 2
    [Abstract] RNA-protein interactions play a crucial role in every aspect of RNA metabolism, and also plays a major role in post-transcriptional gene regulation. RNA-binding proteins have been implicated in viral gene expression (Ray and Das, 2002) and microRNA-mediated gene regulation (Poria et al., 2016). Here we have described the protocol which ...
    Polysome Analysis
    Authors:  Dipak Kumar Poria and Partho Sarothi Ray, date: 03/20/2017, view: 8777, Q&A: 1
    [Abstract] Polysome analysis is a method to separate mRNAs from a cell into actively translating and non-translating fractions depending on their association with polysomes. By this protocol, cell lysates are fractionated by sucrose density gradient ultracentrifugation. Free mRNA fraction and various ribosomal fractions, such as 40S, 60S, monosomes and ...



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