Cancer Biology

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    Protocols in Current Issue
    Measuring Protein Synthesis during Cell Cycle by Azidohomoalanine (AHA) Labeling and Flow Cytometric Analysis
    Authors:  Koshi Imami and Tomoharu Yasuda, date: 04/20/2019, view: 554, Q&A: 0
    [Abstract] Protein synthesis is one of the most fundamental biological processes to maintain cellular proteostasis. Azidohomoalaine (AHA) is a non-radioactive and “clickable” amino acid analog of methionine which can be incorporated into newly synthesized proteins. Thus, AHA-labeled nascent proteins can be detected and quantified through fluorescent labeling ...
    Hyaluronan Isolation from Mouse Mammary Gland
    Authors:  Cornelia Tolg, Mary Cowman and Eva A. Turley, date: 06/05/2018, view: 1359, Q&A: 0
    [Abstract] The glycosaminoglycan hyaluronan (HA) is a key component of the extracellular matrix. The molecular weight of HA is heterogeneous and can reach from several million to several hundred daltons. The effect of HA on cell behavior is size dependent; fragmented HA acts as a danger signal, stimulates cell migration and proliferation and is ...
    Analyzing the Properties of Murine Intestinal Mucins by Electrophoresis and Histology
    Authors:  Ran Wang and Sumaira Z. Hasnain, date: 07/20/2017, view: 8281, Q&A: 0
    [Abstract] Specialized secretory cells known as goblet cells in the intestine and respiratory epithelium are responsible for the secretion of mucins. Mucins are large heavily glycosylated proteins and typically have a molecular mass higher than 106 Da. These large proteins are densely substituted with short glycan chains, which have many important ...
    Single-molecule Analysis of DNA Replication Dynamics in Budding Yeast and Human Cells by DNA Combing
    Authors:  Hélène Tourrière, Julie Saksouk, Armelle Lengronne and Philippe Pasero , date: 06/05/2017, view: 5974, Q&A: 0
    [Abstract] The DNA combing method allows the analysis of DNA replication at the level of individual DNA molecules stretched along silane-coated glass coverslips. Before DNA extraction, ongoing DNA synthesis is labeled with halogenated analogues of thymidine. Replication tracks are visualized by immunofluorescence using specific antibodies. Unlike biochemical ...
    Nucleosome Positioning Assay
    Authors:  Zhongliang Zhao and Holger Bierhoff, date: 05/20/2017, view: 5316, Q&A: 0
    [Abstract] The basic unit of chromatin is the nucleosome, a histone octamer with 147 base pairs of DNA wrapped around it. Positions of nucleosomes relative to each other and to DNA elements have a strong impact on chromatin structure and gene activity and are tightly regulated at multiple levels, i.e., DNA sequence, transcription factor binding, ...
    Liposome Flotation Assays for Phosphoinositide-protein Interaction
    Authors:  Helene Tronchere and Frederic Boal, date: 03/05/2017, view: 8187, Q&A: 0
    [Abstract] Phosphoinositides are rare membrane lipids involved in the control of the major cellular functions and signaling pathways. They are able to recruit specific effector proteins to the cytosolic face of plasma membrane and organelles to coordinate a vast variety of signaling and trafficking processes, as well to maintain specific identity of the ...
    In situ Hybridization (ISH) and Quantum Dots (QD) of miRNAs
    Authors:  Sajni Josson, Murali Gururajan and Leland W.K. Chung, date: 02/20/2017, view: 5323, Q&A: 0
    [Abstract] miRNA are short non-coding RNA which inhibit translation of mRNA. miRNA regulate several cellular processes. Certain miRNA are known to induce oncogenesis. miRNA can be measured by real-time PCR and be imaged using a combination of in situ hybridization (ISH) and quantum dots (QD). The advantage of using quantum dots is that several miRNA ...
    miRNA Characterization from the Extracellular Vesicles
    Authors:  Sajni Josson, Murali Gururajan and Leland W.K. Chung, date: 02/20/2017, view: 3692, Q&A: 0
    [Abstract] Cancer cells and cancer associated stromal cells co-evolve secrete extracellular vesicles to the surrounding regions and regulate several processes involved in cancer metastasis. miRNAs have been known to be mediators of cancer progression and metastasis. miRNAs consist of short noncoding RNA. miRNAs are stable in extracellular fluids such as ...
    Polysome Fractionation to Analyze mRNA Distribution Profiles
    Authors:  Amaresh C. Panda, Jennifer L. Martindale and Myriam Gorospe, date: 02/05/2017, view: 11499, Q&A: 2
    [Abstract] Eukaryotic cells adapt to changes in external or internal signals by precisely modulating the expression of specific gene products. The expression of protein-coding genes is controlled at the transcriptional and post-transcriptional levels. Among the latter steps, the regulation of translation is particularly important in cellular processes that ...
    In vitro Assessment of RNA Polymerase I Activity
    Author:  Marzia Govoni, date: 02/05/2017, view: 3868, Q&A: 0
    [Abstract] In eukaryotic cells transcriptional processes are carried out by three different RNA polymerases: RNA polymerase I which specifically transcribes ribosomal RNA (rRNA), RNA polymerase II which transcribes protein-coding genes to yield messenger RNAs (mRNAs) and small RNAs, while RNA polymerase III transcribes the genes for transfer RNAs and for the ...



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