Cancer Biology

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    Protocols in Current Issue
    Automated Analysis of Cell Surface Ruffling: Ruffle Quantification Macro
    Authors:  Nicholas D. Condon, Jennifer L. Stow and Adam A. Wall, date: 01/20/2020, view: 585, Q&A: 1
    [Abstract] Cell surface protrusions include F-actin rich, wave-like ruffles that are erected transiently in response to stimuli and during cell migration. Macrophages are innate immune cells that ruffle constitutively and more dramatically in cells activated by pathogens. Dorsal ruffles and their resulting macropinosomes are key sites for environmental ...
    Immunofluorescence-based Determination of Centrosome Number in Tissue Samples
    Authors:  Mengdie Wang, Gregory C. Rogers and Anne E. Cress, date: 10/20/2019, view: 1403, Q&A: 0
    [Abstract] Centrosome numerical abnormalities have been reported in a variety of tumors. Centrosome numbers in cancer cells display both inter-tumor and intra-tumor heterogeneity. The over production of centrosomes (centrosome amplification) is unique in cancer cells and is a promising target for therapy. Thus, a method to quantify centrosome numbers on a ...
    RNA Interference Screening to Identify Proliferation Determinants in Breast Cancer Cells
    [Abstract] RNAi screening technology has revealed unknown determinants of various biological signaling pathways in biomedical studies. This protocol provided detailed information about how to use RNAi screening to identify proliferation determinants in breast tumor cells. siRNA-based libraries targeting against Estrogen receptor (ER)-network, including 631 ...
    Measurement of the Intracellular Calcium Concentration with Fura-2 AM Using a Fluorescence Plate Reader
    Authors:  Magdiel Martínez, Namyr A. Martínez and Walter I. Silva, date: 07/20/2017, view: 19896, Q&A: 1
    [Abstract] Intracellular calcium elevation triggers a wide range of cellular responses. Calcium responses can be affected or modulated by membrane receptors mutations, localization, exposure to agonists/antagonists, among others (Burgos et al., 2007; Martínez et al., 2016). Changes in intracellular calcium concentration can be measured ...
    Soft Agar Colony Formation Assay as a Hallmark of Carcinogenesis
    Authors:  Feng Du, Xiaodi Zhao and Daiming Fan, date: 06/20/2017, view: 18422, Q&A: 0
    [Abstract] Soft agar colony formation assay is established to estimate the anchorage-independent growth ability of cells. In this assay, a bottom layer of agar with complete media is poured and solidified first, followed by an upper layer containing a specified number of cells suspended in medium-agar mixture. After two weeks of incubation, the number of ...
    Isolation of Murine Alveolar Type II Epithelial Cells
    Authors:  Fan Sun, Gutian Xiao and Zhaoxia Qu, date: 05/20/2017, view: 6895, Q&A: 0
    [Abstract] We have optimized a protocol for isolation of alveolar type II epithelial cells from mouse lung. Lung cell suspensions are prepared by intratracheal instillation of dispase and agarose followed by mechanical disaggregation of the lungs. Alveolar type II epithelial cells are purified from these lung cell suspensions through magnetic-based negative ...
    Murine Bronchoalveolar Lavage
    Authors:  Fan Sun, Gutian Xiao and Zhaoxia Qu, date: 05/20/2017, view: 15842, Q&A: 1
    [Abstract] A basic Bronchoalveolar lavage (BAL) procedure in mouse is described here. Cells and fluids obtained from BAL can be analyzed by Hema3-staining, immunostaining, Fluorescence-activated cell sorting (FACS), PCR, bicinchoninic acid protein assay, enzyme-linked immunosorbent assay (ELISA), luminex assays, etc., to examine the immune cells, ...
    Semi-quantitative Analysis of H4K20me1 Levels in Living Cells Using Mintbody
    Authors:  Yuko Sato and Hiroshi Kimura, date: 05/20/2017, view: 5623, Q&A: 0
    [Abstract] Eukaryotic nuclear DNA wraps around histone proteins to form a nucleosome, a basic unit of chromatin. Posttranslational modification of histones plays an important role in gene regulation and chromosome duplication. Some modifications are quite stable to be an epigenetic memory, and others exhibit rapid turnover or fluctuate during the cell cycle. ...
    Immunostaining of Formaldehyde-fixed Metaphase Chromosome from Untreated and Aphidicolin-treated DT40 Cells
    Author:  Vibe H. Oestergaard, date: 05/05/2017, view: 6286, Q&A: 0
    [Abstract] During mitosis chromosomes are condensed into dense X-shaped structures that allow for microscopic determination of karyotype as well as inspection of chromosome morphology.

    This protocol describes a method to perform immunostaining of formaldehyde-fixed metaphase chromosomes from the avian cell line DT40. It was developed to ...
    Preparation of Primary Astrocyte Culture Derived from Human Glioblastoma Multiforme Specimen
    Authors:  Mansoureh Hashemi and Mahmoudreza Hadjighassem, date: 04/20/2017, view: 6735, Q&A: 0
    [Abstract] Glioblastoma multiforme (GBM) is a grade 4 astrocytoma tumor in central nervous system. Astrocytes can be isolated from human GBM. Study of astrocytes can provide insights about the formation, progression and recurrence of glioblastoma. For using isolated astrocytes, new studies can be designed in the fields of pharmacology, neuroscience and ...



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