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

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.

Get tips on using EGMTM-2 Endothelial Cell Growth Medium-2 BulletKit to perform Stem cell culture media hLung Mesenchymal Stem Cells (MSCs)

Get tips on using Fenozol to perform siRNA / miRNA gene silencing Human - HeLa Cdc20

Get tips on using SIHK1738 to perform siRNA / miRNA gene silencing Human - LLC PKN3

Get tips on using SIHK1738 to perform siRNA / miRNA gene silencing Human - HeLa PKN3

Get tips on using NM_001350 to perform siRNA / miRNA gene silencing Human - U2OS DAXX

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary human pulmonary artery endothelial cells

Get tips on using TaqMan® MicroRNA Reverse Transcription Kit to perform siRNA / miRNA gene silencing Mouse - Glomerular mesangial cells HIPK2 Polymer / Lipid delivery

Get tips on using FGD4 siRNAs (FlexiTube) to perform siRNA / miRNA gene silencing Human - LNCap FGD4

Get tips on using EGMTM-2 Endothelial Cell Growth Medium-2 BulletKit to perform Mammalian cell culture media RAOEC