ChIP H3K36Me3 Sheep Rat

Get tips on using ChIP-IT® Express Chromatin Immunoprecipitation Kits to perform ChIP Rat - H4IIE

Get tips on using Magna ChIP™ A - Chromatin Immunoprecipitation Kit to perform ChIP Rat - Liver

Get tips on using Magna ChIP™ A - Chromatin Immunoprecipitation Kit to perform ChIP Rat - Brain

Get tips on using ChIP-IT® Express Chromatin Immunoprecipitation Kits to perform ChIP Rat - Mesenteric arteries

Get tips on using ChIP-IT® Express Chromatin Immunoprecipitation Kits to perform ChIP Rat - INS-1

Get tips on using EZ-Magna ChIP™ G - Chromatin Immunoprecipitation Kit to perform ChIP Rat - Liver

Get tips on using SOLiD™ ChIP-Seq Kit, with ChIP magnet to perform ChIP Human - SH-SY5Y

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.

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 in the experimental, add more DNA to the PCR reaction or increase the number of amplification cycles. Choose an alternate, ChIP-validated antibody if the antibody does not work.

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.