Single-cell qPCR Assay with Massively Parallel Microfluidic System
[Abstract] The single-cell transcriptome is the set of messenger RNA molecules expressed in one cell. It is extremely variable and changes according to external, physical and biochemical conditions. Due to sensitivity shortages, most of genetic studies use bulk samples, providing only the average gene expression. Single-cell technologies have provided a ...
Rapid Nickel-based Isolation of Extracellular Vesicles from Different Biological Fluids
Authors: Michela Notarangelo,
Deborah Ferrara,
Cristina Potrich,
Lorenzo Lunelli,
Lia Vanzetti,
Alessandro Provenzani,
Manuela Basso,
Alessandro Quattrone and
Vito G. D'Agostino,
date: 02/05/2020,
view: 653,
Q&A: 0
[Abstract] Extracellular vesicles (EVs) are membranous structures that cells massively release in extracellular fluids. EVs are cargo of cellular components such as lipids, proteins, and nucleic acids that can work as a formidable source in liquid biopsy studies searching for disease biomarkers. We recently demonstrated that nickel-based isolation (NBI) is a ...
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. ...
Approaching RNA-seq for Cell Line Identification
Authors: Tabrez A. Mohammad and
Yidong Chen,
date: 02/05/2020,
view: 502,
Q&A: 0
[Abstract] Cancer cell lines serve as invaluable model systems for cancer biology research and help in evaluating the efficacy of new therapeutic agents. However, cell line contamination and misidentification have become one of the most pressing problems affecting biomedical research. Available methods of cell line authentication suffer from limited access, ...
Bone-in-culture Array to Model Bone Metastasis in ex vivo Condition
Authors: Hai Wang and
Xiang H.-F. Zhang,
date: 01/20/2020,
view: 472,
Q&A: 0
[Abstract] Bone is the most frequently affected organ by metastases of breast cancer and prostate cancer. Our knowledge on bone metastasis is extremely limited due to the lack of potent and efficient experimental models. We developed the “Bone-In-Culture Array (BICA)” platform to model the bone colonization of cancer cells in ex vivo cultures. The ...
Automated Analysis of Cell Surface Ruffling: Ruffle Quantification Macro
[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 ...
Analysis of Random Migration of Cancer Cells in 3D
Authors: Sai P. Visweshwaran and
Alexis Gautreau,
date: 01/05/2020,
view: 631,
Q&A: 0
[Abstract] The ability of cancer cells to migrate through a complex three-dimensional (3D) environment is a hallmark event of cancer metastasis. Therefore, an in vitro migration assay to evaluate cancer cell migration in a 3D setting is valuable to examine cancer progression. Here, we describe such a simple migration assay in a 3D ...