Cancer Biology

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    A mant-GDP Dissociation Assay to Compare the Guanine Nucleotide Binding Preference of Small GTPases
    Authors:  Yuping Tan and Qingxiang Sun, date: 01/20/2021, view: 1224, Q&A: 0

    Small GTPases are cellular switches that are switched on when bound to GTP and switched off when bound to GDP. Different small GTPase proteins or those with mutations may bind to GTP or GDP with different relative affinities. However, small GTPases

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    Real-time Three-dimensional Tracking of Endocytic Vesicles
    Authors:  Stephanie Duhamel and Kossay Zaoui, date: 10/20/2020, view: 1338, Q&A: 0
    Endocytic trafficking and recycling are fundamental cellular processes that control essential functions such as signaling protein complexes transport and membrane identity. The small GTPase Rabs are indispensable component of the endosomal recycling ...
    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: 1522, Q&A: 0
    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 ...
    Colorimetric RhoB GTPase Activity Assay
    Authors:  Kossay Zaoui and Stephanie Duhamel, date: 05/05/2020, view: 2274, Q&A: 0
    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 ...
    Identification of Target Protein for Bio-active Small Molecule Using Photo-cross Linked Beads and MALDI-TOF Mass Spectrometry
    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 ...
    Measurement of ATPase Activity of Valosin-containing Protein/p97
    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 ...
    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: 3544, Q&A: 0
    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 ...
    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: 5782, Q&A: 0
    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 ...
    Purifying Properly Folded Cysteine-rich, Zinc Finger Containing Recombinant Proteins for Structural Drug Targeting Studies: the CH1 Domain of p300 as a Case Example
    Authors:  Yong Joon Kim, Stefan Kaluz, Anil Mehta, Emily Weinert, Shannon Rivera and Erwin G. Van Meir, date: 09/05/2017, view: 10391, Q&A: 0
    The transcription factor Hypoxia-Inducible Factor (HIF) complexes with the coactivator p300, activating the hypoxia response pathway and allowing tumors to grow. The CH1 and CAD domains of each respective protein form the interface between p300 and ...
    Non-radioactive LATS in vitro Kinase Assay
    Authors:  Audrey W. Hong and Kun-Liang Guan, date: 07/20/2017, view: 9050, Q&A: 1
    This protocol describes a method to directly measure LATS activity by an in vitro kinase assay using YAP as a substrate.



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