ChIP H3K27me3 Human Sheep

In ChIP, the most vital step is the binding of an antibody and choosing the right antibody. The binding affinity of different types of immunoglobulins to protein A or G differs significantly. Henceforth, it is recommended to choose either protein A or protein G coated beads. If you do not see any product in the positive control, add 5–10 μg of chromatin and 1–5 μg of antibody to each IP reaction and incubate with antibody overnight and an additional 2 hr after adding Protein G/A beads. If no product is observed in the experimental sample, add more DNA to the PCR reaction or increase the number of amplification cycles. Furthermore, if you have any problem with antibodies, make sure to use the ChIP-validated antibody.

Get tips on using Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade to perform ChIP H3K27me3 - Sheep Rat YFP Tag

Get tips on using Anti-trimethyl-Histone H3 (Lys27) Antibody to perform ChIP H3K27me3 - Sheep Rat YFP Tag

Get tips on using Anti-Histone H3 (acetyl K27) antibody - ChIP Grade (ab4729) to perform ChIP Anti-bodies H3K27me3

Get tips on using Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade to perform ChIP H3K36Me3 - Sheep Rat -NA-

Get tips on using Histone H3K9ac antibody to perform ChIP H3K9Ac - Sheep Rat -NA-

Get tips on using Anti-trimethyl-Histone H3 (Lys27) Antibody to perform ChIP Anti-bodies H3K27me3

Get tips on using Anti-trimethyl-Histone H3 (Lys9) Antibody to perform ChIP H3K9me3 - Sheep Rat -NA-

Get tips on using Anti-trimethyl-Histone H3 (Lys4) Antibody to perform ChIP H3K4Me3 - Sheep Rat -NA-

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.