Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - immortalized CMT12
Wound healing assay can be challenging due to inconsistencies and variations while making a wound on the confluent cell monolayer, consequently leads to wounds of varying sizes and widths. Moreover, this assay causes damage to the cells that are at the edge of the wound, which can prevent cell migration into the wound site and healing. The best solution is to use the standard wound healing assay kits using either combs or inserts to make a defined wound field or gap and prevent the well-to-well variation in these assays.
Reporter gene assays are designed to test the regulation of the expression of a gene of interest. This is usually done by linking the promoter of the gene of interest with a gene such as a firefly luciferase, which can be easily detected by addition of luciferin that leads to an enzymatic reaction to produce luminescence. The enzymatic reaction can be correlated to the expression of the gene of interest. Another luciferase gene that can be used is Renilla luciferase. For an appropriate luciferase assay: 1. the reporter should express uniformly in all cells, 2. specifically respond to effectors that the assay intends to monitor, 3. have low intrinsic stability to quickly reflect transcriptional dynamics. It is important to have an equal number of cells plated in each testing condition to avoid any incorrect readouts. Reporter assays could be single or dual reporter assays. The reporter could be both luciferases. Most dual-luciferase assays involve adding two reagents to each sample and measuring luminescence following each addition. Adding the first reagent activates the first luciferase reporter reaction; adding the second reagent extinguishes first luciferase reporter activity and initiates the second luciferase reaction. Dual-luciferase assays have some advantages, including 1. reduces variability, 2. reduces background, 3. normalizes differences in transfection efficiencies between samples.
Get tips on using GeneChip® Human Genome U133 Plus 2.0 Array to perform RNA amplification & labeling Mammalian - RNA, rhesus monkey brain tissue Human endothelail stromal cells
Get tips on using QIAamp DNA Mini Kit to perform DNA isolation / purification Cells - Immortalized cell lines Renal cortical tubule epithelial cells
Get tips on using SV Total RNA Isolation System to perform RNA isolation / purification Cells - primary human coronary artery smooth muscle cells
Get tips on using PureLink™ RNA Mini Kit to perform RNA isolation / purification Cells - primary canine coronary artery smooth muscle 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
Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary human airway epithelial cells
Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary human aortic endothelial cells
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