Autophagy assay cell type

Western blotting is a widely used technique to size separate proteins from a pool of cell or tissue lysates. The technique has 4 major steps: a) gel electrophoresis, b) blocking and treatment with antigen specific antibody, c) treatment with secondary antibody and finally d) detection and visualization. Though western blotting is a widely used technique, detection of specific proteins depends on several factors, the major ones are antibody concentration, incubation time and washing steps. Key points for obtaining clean blots are: always prepare fresh buffer solutions and optimize antibody concentration. Given the advent of high-throughput protein analysis and a push to limit the use of lab consumables, onestep antibodies are developed which recognise protein of interest and also contain a detection label.

Get tips on using Image-IT™ LIVE Green Reactive Oxygen Species Detection Kit, for microscopy to perform ROS assay cell type - H9c2 rat cardiomyocytes

Get tips on using ROS-ID® Total ROS/Superoxide detection kit to perform ROS assay cell type - A549 human adenocarcinomic human alveolar basal epithelial cells

Get tips on using ApopTag® Fluorescein In Situ Apoptosis Detection Kit to perform TUNEL assay cell type - A549, NCI-H460, H1299 human lung cancer cells

Get tips on using FragEL™ DNA Fragmentation Detection Kit, Colorimetric - TdT Enzyme to perform TUNEL assay cell type - A127, U87MG, U251MG, T98G human glioblastoma cells

A standard angiogenic assay involves the autonomous endothelial cell response of self-organization into microvessels, also known as tubes when seeded on a basement membrane matrix in the presence of the appropriate growth factors. However, the component of basement membrane matrix may also affect the tube formation by endothelial cells. Hence it is important to use a standard angiogenesis assay kit or use the same membrane matrix with known composition to standardize the assay conditions.

Get tips on using CM-H2DCFDA (General Oxidative Stress Indicator) to perform ROS assay cell type - PLHC-1, SK-HEP-1, Hep3b, HepG2 human hepatocellular carcinoma

Get tips on using Carboxy-H2DCFDA (general oxidative stress indicator) to perform ROS assay cell type - PLHC-1, SK-HEP-1, Hep3b, HepG2 human hepatocellular carcinoma

Get tips on using ApopTag® Peroxidase In Situ Apoptosis Detection Kit to perform TUNEL assay cell type - HNSCC Detroit 562 human head and neck tumor 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.