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.
Get tips on using CD80 (B7-1) Monoclonal Antibody (16-10A1), APC, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD80
Get tips on using CD80 (B7-1) Monoclonal Antibody (16-10A1), PE, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD80
When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.
Get tips on using Color-coded Prestained Protein Marker, Low Range (1.7-42 kDa) #13070 to perform Protein Ladder Prestained
Get tips on using Prestained Protein Ladder – Mid-range molecular weight (10 - 180 kDa) (ab116027) to perform Protein Ladder Prestained
Get tips on using Blu13 (BLUelf) Prestained Protein Ladder（5 to 245 kDa） to perform Protein Ladder Prestained
Get tips on using Color-coded Prestained Protein Marker, High Range (43-315 kDa) #12949 to perform Protein Ladder Prestained
Get tips on using Color-coded Prestained Protein Marker, Broad Range (10-250 kDa) #74124 to perform Protein Ladder Prestained
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