Cancer Biology

Categories

    Protocols in Current Issue
    Tyramide Signal-Amplified Immunofluorescence of MYCN and MYC in Human Tissue Specimens and Cell Line Cultures
    Authors:  Johanna M. Schafer and Jennifer A. Pietenpol, date: 07/05/2020, view: 554, Q&A: 0
    [Abstract] MYC family members, MYC, MYCN, and MYCL, are oncogenic transcription factors that regulate the expression of genes involved in normal development, cell growth, proliferation, metabolism, and survival. While MYC is amplified and/or overexpressed across a variety of tissue types, MYCN is often overexpressed in tumors of the nervous system ...
    Colorimetric RhoB GTPase Activity Assay
    Authors:  Kossay Zaoui and Stephanie Duhamel, date: 05/05/2020, view: 1271, Q&A: 0
    [Abstract] The Ras homologous protein (Rho) GTPase subfamily, including RhoA, RhoB, and RhoC are small molecules (~21 kDa) that act as molecular switches in a wide range of signaling pathways to orchestrate biological processes associated with both physiological and tumorigenic cellular states. The Rho GTPases are crucial regulators of actin cytoskeleton ...
    Identification of Target Protein for Bio-active Small Molecule Using Photo-cross Linked Beads and MALDI-TOF Mass Spectrometry
    [Abstract] Development of methods for protein identification is one of the important aspects of proteomics. Here, we report a protocol for the preparation of compound conjugated beads by photo-crosslinking, affinity purification, gel electrophoresis, and highly sensitive mass spectrometric assay for drug-target identification. Although there are several ...
    Measurement of ATPase Activity of Valosin-containing Protein/p97
    [Abstract] Valosin-containing protein (VCP; also known as p97) is a type II ATPase regulating several cellular processes. Using proteomic techniques, we identified a chemical compound that binds to the D1 ATPase domain of VCP. The protocol described here was to study the effect of the compound on ATPase activity in vitro of purified VCP protein. ...
    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: 1835, 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: 4856, 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: 7716, 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: 4052, 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: 7636, 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: 6646, 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, ...



    We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.