Immunohistochemistry Collagen VII [II-32]

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.

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.

Isolating RNA from tissues and paraffin-embedded tissue samples can be challenging due to cross-linking of biomolecules and fragmented nucleic acids. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in presence of RNAse inhibitors. The homogenization process should be carried out on dry ice to maintain the integrity of RNA.

Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Tissue - Human Veins

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human umbilical vein smooth muscle cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human umbilical vein endothelial cells

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human umbilical vein smooth muscle cells

Get tips on using TRI Reagent® MRC to perform RNA isolation / purification Cells - primary human umbilical vein endothelial cells

Get tips on using PureLink™ RNA Mini Kit to perform RNA isolation / purification Cells - primary human umbilical vein endothelial cells

Get tips on using E.Z.N.A.® Total RNA Kit I to perform RNA isolation / purification Cells - primary bovine umbilical vein endothelial cells