ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.
Cell cycle can be challenging due to difference introduced by sample handling, timing, and difference within the sample. Downstream instriuments to analyse cell cycle (Multicolor flow cytometry and multicolor imaging) can answer these challenges. Relevant markers can be combined with cell phenotyping markers to look at events within subpopulations of cells.
Get tips on using DeadEnd™ Fluorometric TUNEL System to perform TUNEL assay cell type - SKOV3, Caov3 human ovarian cancer
Get tips on using MEBSTAIN Apoptosis TUNEL Kit Direct to perform TUNEL assay cell type - PANC-1 human pancriatic cancer
Get tips on using Senescence β-Galactosidase Staining Kit - Beyotime to perform Reporter gene assay β-galactosidase substrates - HeLa cervical cancer cells
Get tips on using β-Galactosidase Reporter Gene Staining Kit to perform Reporter gene assay β-galactosidase substrates - HeLa cervical cancer cells
Get tips on using Luminescent β-galactosidase Detection Kit II to perform Reporter gene assay β-galactosidase substrates - HeLa cervical cancer cells
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.
Get tips on using TUNEL Assay Kit - BrdU-Red to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells
An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.
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