siRNA / miRNA gene silencing Rat Retinal stem cells

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Get tips on using MagNA Pure Compact RNA Isolation Kit to perform RNA isolation / purification Cells - primary human mesenchymal stem cells

Products Roche Lifesciences MagNA Pure Compact RNA Isolation Kit

Get tips on using Pierce™ Coomassie (Bradford) Protein Assay Kit to perform Protein quantification Mammalian cells - Human pluripotent stem cells

Products Thermo Fisher Scientific Pierce™ Coomassie (Bradford) Protein Assay Kit

Get tips on using GATA-1 shRNA Plasmids (h) to perform shRNA gene silencing Human - TF‐1 GATA‐1

Products Santa Cruz Biotechnology GATA-1 shRNA Plasmids (h)

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.

Cellular assays Reporter gene assay β-galactosidase substrates SK-Hep-1

Get tips on using Iscove’s Modified Dulbecco’s Medium to perform Stem cell culture media hMegakaryoblastic cells

Products Sigma-Aldrich Iscove’s Modified Dulbecco’s Medium

Get tips on using Rock-2 siRNA and shRNA Plasmids (h) to perform RNA sequencing Human - HT-1376 (urinary bladder cell line)

Products Santa Cruz Biotechnology Rock-2 siRNA and shRNA Plasmids (h)

Get tips on using β-Gal Reporter Gene Assay, chemiluminescent to perform Reporter gene assay β-galactosidase substrates - RAW 264.7

Products Sigma-Aldrich β-Gal Reporter Gene Assay, chemiluminescent

Get tips on using StemMACS™ AdipoDiff Media, human to perform Stem cell Differentiation media hMSCs differentiation into adipogenic cells

Products Miltenyibiotec StemMACS™ AdipoDiff Media, human

Get tips on using FuGENE® 6 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat hepatic stellate cells

Products Promega FuGENE® 6 Transfection Reagent

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat hepatic stellate cells

Products Thermo Fisher Scientific Lipofectamine® 2000 Transfection Reagent

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