Get tips on using DMEM/F-12, no phenol red to perform 3D Cell Culture Media Human breast cancer MDA-MB-231 cells-Mammospheres
Get tips on using DMEM/F-12 PLUS Basal Medium to perform 3D Cell Culture Media Human breast cancer MDA-MB-231 cells-Mammospheres
Get tips on using Gibco™ DMEM, high glucose, GlutaMAX™ Supplement to perform Stem cell culture media Human Dental pulp stem cells (hDPSC)
Get tips on using QIA33 | FragEL™ DNA Fragmentation Detection Kit, Colorimetric - TdT Enzyme to perform Apoptosis assay cell type - Human endometrial stromal cells
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
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
Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.
As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.
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
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