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.
Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase
Wound healing assay can be challenging due to inconsistencies and variations while making a wound on the confluent cell monolayer, consequently leads to wounds of varying sizes and widths. Moreover, this assay causes damage to the cells that are at the edge of the wound, which can prevent cell migration into the wound site and healing. The best solution is to use the standard wound healing assay kits using either combs or inserts to make a defined wound field or gap and prevent the well-to-well variation in these assays.
Get tips on using In Situ Cell Death Detection Kit, Fluorescein to perform TUNEL assay cell type - A549, NCI-H460, H1299 human lung cancer cells
Get tips on using in situ Cell Death Detection Kit, POD to perform TUNEL assay cell type - A549, NCI-H460, H1299 human lung cancer cells
Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - immortalized HeLa, CaSki and C33A (Cervical cancer cells)
Get tips on using Click-iT™ TUNEL Alexa Fluor™ 488 Imaging Assay to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells
A gross majority of classical apoptotic attributes can be quantitatively examined by flow cytometry, the preferred platform for rapid assessment of multiple cellular attributes at a single-cell level. However, sample 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.
Get tips on using In Situ Cell Death Detection Kit, TMR red to perform TUNEL assay cell type - A549, NCI-H460, H1299 human lung cancer cells
Get tips on using DeadEnd™ Fluorometric TUNEL System to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells
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