ChIP Rat - Colon

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.

Start discussion

No discussions found

Share your thoughts or question with experts in your field

Start a discussion

Found 1 matching solution for this experiment

EpiQuik Chromatin Immunoprecipitation (ChIP) Kit

Epigentek

Upstream tips	Protocol tips	Downstream tips
-Ensure that all buffers are in clear solution. Shake or vortex if these buffers precipitate.	-When processing spin columns, always cap spin columns before placing them in the microcentrifuge. -The conditions of crosslinked DNA shearing can be optimized based on cells and sonicator equipment. If desired, remove 5 µl of sonicated cell lysate for agarose gel analysis. The length of sheared DNA should be between 200-1000 bp.

Upstream tips
-Ensure that all buffers are in clear solution. Shake or vortex if these buffers precipitate.
Protocol tips
-When processing spin columns, always cap spin columns before placing them in the microcentrifuge. -The conditions of crosslinked DNA shearing can be optimized based on cells and sonicator equipment. If desired, remove 5 µl of sonicated cell lysate for agarose gel analysis. The length of sheared DNA should be between 200-1000 bp.

Manufacturer protocol Publication protocol 1 Relevant paper

Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product!

Fill out your contact details and receive price quotes in your Inbox