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 Corning® BioCoat™ Matrigel® Invasion Chamber with 8.0 µm PET Membrane in four 6-well Plates to perform Cell migration / Invasion cell type - 4T1
Get tips on using Corning® BioCoat™ Matrigel® Invasion Chambers with 8.0 µm PET Membrane in two 24-well Plates to perform Cell migration / Invasion cell type - BxPC-3
Get tips on using Corning® BioCoat™ Matrigel® Invasion Chambers with 8.0 µm PET Membrane in two 24-well Plates to perform Cell migration / Invasion cell type - MG-63
The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.
Get tips on using DMEM to perform Mammalian cell culture media HFLS
Get tips on using IMDM to perform Mammalian cell culture media KG1
Get tips on using IMDM to perform Mammalian cell culture media U937
Get tips on using Dansylcadaverine to perform Autophagy assay cell type - C6
Get tips on using Dansylcadaverine to perform Autophagy assay cell type - MRC5
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