Site Directed Mutagenesis (SDM) Human Deletion MDA-MB-231

Get tips on using The Premo Autophagy Tandem Sensor RFP-GFP-LC3B Kit to perform Autophagy assay cell type - Human osteosarcoma cancer cells

Get tips on using CD36 Monoclonal Antibody (eBioNL07 (NL07)), PerCP-eFluor 710, eBioscience™ to perform Flow cytometry Anti-bodies Human - CD36/CB38

Get tips on using Ambion™ RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE to perform RNA isolation / purification Tissue - Human Kidney

Get tips on using Click-iT™ EdU Pacific Blue™ Flow Cytometry Assay Kit to perform Cell cycle assay human - MCF 10A

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

Get tips on using Click-iT™ Plus EdU Pacific Blue™ Flow Cytometry Assay Kit to perform Cell cycle assay human - A549

Get tips on using Click-iT™ Plus EdU Pacific Blue™ Flow Cytometry Assay Kit to perform Cell cycle assay human - SKOV3

Cell cytotoxicity assays measure the ability of certain compounds or chemical mediators to reduce the viability of the cells. The term cell cytotoxicity assay can sometimes be used interchangeably with cell proliferation assay. Healthy living cells can be identified by the use of formazan dyes, protease biomarkers or by measuring ATP content. The formazan dyes are chromogenic products formed by the reduction of tetrazolium salts by dehydrogenases, such as lactate dehydrogenase (LDH) and reductases that are released during cell death. Common tetrazolium salts include INT, MTT, MTS and XTT. Cell cytotoxicity can also be measured by using the SRB and WST-1 assays. These assays can usually be used in a high-throughput fashion and can be quantitated by measuring absorbance, colorimetry or luminescence. All these assays require similar numbers of cell plating at the initiation, a time course of treatment with the cytotoxic agent and at least triplicates for each condition at every point of analysis. Cell shrinkage, plasma membrane blebbing, cell detachment, externalization of phosphatidylserine, nuclear condensation and ultimately DNA fragmentation are well-described features of apoptosis. The assays that rely on cell membrane integrity for their function, may not be able to quantify early apoptosis. Therefore, in order to distinguish early apoptotic vs. late apoptotic or necrotic cells, additional flow cytometry techniques can be used. A combination of Annexin V and PI (propidium iodide) can be used to distinguish early (Annexin V+/PI-) and late apoptotic (Annexin V+/PI+) cells. Sometimes, caspase assays are used in order to differentiate the stages of apoptosis.

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.

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.