siRNA / miRNA gene silencing Human HUVEC

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RNA siRNA / miRNA gene silencing Rat Glial cells C/EBP‐β

RNA siRNA / miRNA gene silencing Rat Retinal stem cells Brn-3b

RNA siRNA / miRNA gene silencing Rat Brain endothelial cells HIF-1α Lipid

RNA siRNA / miRNA gene silencing Mouse Glomerular mesangial cells HIPK2 Polymer / Lipid delivery

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

Cellular assays Reporter gene assay β-galactosidase substrates HUVEC

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DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

Proteins ChIP Human HUVEC

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Get tips on using siGENOME Human MAP4K2 siRNA to perform siRNA / miRNA gene silencing Human - RMS MAP4K2

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