tunel-assay-cell-type-panc-1-human-pancriatic-cancer

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As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.

Cellular assays Autophagy assay cell type U2OS (human bone osteosarcoma epithelial cells)

Cell culture media 3D Cell Culture Media Human breast cancer MCF-7 cells-Mammospheres

Cell Invasion or Cell Migration assays are technically challenging to set up as the gradient between the two compartments equilibrates in time during the assay. It is also problematic to view cells and for cells to migrate through a non-physiologic polycarbonate or polypropylene filter. Care must be taken while loading the well with cells to form a single cell suspension. Precaution must be taken while trypsinization (under-trypsinization can lead to cell clumping while over-trypsinization could strip off adhesion molecules necessary for migration). This leads to difficulty in getting significant results, when only small numbers of cells cross the filter or when the distribution and/or staining of the cells is uneven.

Cellular assays Cell migration / Invasion cell type LP-1

Get tips on using Click-iT™ EdU Pacific Blue™ Flow Cytometry Assay Kit to perform Cell cycle assay human - PANC-1

Products Thermo Fisher Scientific Click-iT™ EdU Pacific Blue™ Flow Cytometry Assay Kit

Cell Invasion or Cell Migration assays are technically challenging to set up as the gradient between the two compartments equilibrates in time during the assay. It is also problematic to view cells and for cells to migrate through a non-physiologic polycarbonate or polypropylene filter. Care must be taken while loading the well with cells to form a single cell suspension. Precaution must be taken while trypsinization (under-trypsinization can lead to cell clumping while over-trypsinization could strip off adhesion molecules necessary for migration). This leads to difficulty in getting significant results, when only small numbers of cells cross the filter or when the distribution and/or staining of the cells is uneven.

Cellular assays Cell migration / Invasion cell type SK-MEL-1

Cell culture media 3D Cell Culture Media Human breast cancer MDA-MB-231 cells-Mammospheres

Get tips on using WST-1 Cell Proliferation Assay Kit to perform Cell cytotoxicity / Proliferation assay cell type - MG-63

Products Takara Bio Inc WST-1 Cell Proliferation Assay Kit

Get tips on using Anti-LC3B antibody produced in rabbit to perform Autophagy assay cell type - PANC-1

Products Sigma-Aldrich Anti-LC3B antibody produced in rabbit

Get tips on using CYTO-ID® Autophagy detection kit to perform Autophagy assay cell type - PANC-1

Products Enzo Life Sciences CYTO-ID® Autophagy detection kit

Get tips on using OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence) to perform ROS assay cell type - PANC-, BxPC-3 human pancreas

Products Cell Biolabs OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence)

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