Site Directed Mutagenesis (SDM) Human Point mutation Caco-2

Get tips on using Human BMP2 ELISA Kit (ab119581) to perform ELISA Human - BMP-2

Get tips on using Stat3 Antibody (F-2): sc-8019 to perform Western blotting STAT3

Get tips on using E-cadherin monoclonal antibody (ECCD-2) to perform Western blotting E-cadherin

Get tips on using Mouse Lipocalin-2/NGAL DuoSet ELISA to perform ELISA Mouse - NGAL/LCN2

Get tips on using Laminin beta-2/gamma-1 Monoclonal Antibody (A5) to perform Western blotting Laminin subunit Beta-2

Get tips on using Laminin β-2 Antibody (H-1): sc-133241 to perform Western blotting Laminin subunit Beta-2

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.

Get tips on using CD74 Antibody (LN-2): sc-6262 to perform Flow cytometry Anti-bodies Mouse - CD74

A key signature for necrotic cells is the permeabilization of the plasma membrane. Necrosis can be quantified by several cellular and biochemical assays. When studied minutely, it reveals the difficulty in confirmation in secondary induction of necrosis in apoptotic cells. Apoptotic cells are being analyzed to shift to necrotic status owing to membrane permeability at later stages, and thus, discrimination of two cell death becomes critical. Therefore, it is crucial to use a necrosis detection kit or a defined procedure to analyze this unprogrammed form of death in response to immense chemical and physical insults.