Isolation of Tumor Cells Based on Their Distance from Blood Vessels
[Abstract] Differential exposure of tumor cells to microenvironmental cues greatly impacts cell phenotypes, raising a need for position based sorting of tumor cells amenable to multiple OMICs and functional analyses. One such key determinant of tumor heterogeneity in solid tumors is its vasculature. Proximity to blood vessels (BVs) profoundly affects tumor ...
Colorimetric RhoB GTPase Activity Assay
Authors: Kossay Zaoui and
Stephanie Duhamel,
date: 05/05/2020,
view: 1061,
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 ...
Measurement of Protein-Protein Interactions through Microscale Thermophoresis (MST)
[Abstract] The binding interactions of PD-1 and PD-L1 have been studied by surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) over the past few years, but these investigations resulted in controversy regarding the values of the dissociation constant (Kd) (Freeman et al., 2000). MST is a powerful new method for the ...
In vivo Tumor Growth and Spontaneous Metastasis Assays Using A549 Lung Cancer Cells
[Abstract] Metastasis accounts for the majority of cancer related deaths. The genetically engineered mouse (GEM) models and cell line-based subcutaneous and orthotopic mouse xenografts have been developed to study the metastatic process. By using lung cancer cell line A549 as an example, we present a modified protocol to establish the cell line-based ...
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: 1229,
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 ...