Protein expression and purification Mammalian cells HEK 293

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human lung fibroblasts (HLF)

Get tips on using Lipofectamine® LTX Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines HeLa

Get tips on using X-tremeGENE™ HP DNA Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat astrocytes

Get tips on using Live-Dead Cell Staining Kit (BioVision) to perform Live / Dead assay mammalian cells - HepG2

Get tips on using High Pure PCR Product Purification Kit to perform DNA gel extraction / PCR product purification Product size > 15Kb

Get tips on using Live-Dead Cell Staining Kit (BioVision) to perform Live / Dead assay mammalian cells - MDA-MB-231 human breast cancer cells

Get tips on using LIVE/DEAD™ Cell Imaging Kit to perform Live / Dead assay mammalian cells - MDA-MB-231 human breast cancer cells

Get tips on using Agencourt AMPure XP - PCR Purification to perform

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.

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.