ChIP acH3 Bovine Rat

Get tips on using Re-ChIP-IT® to perform ChIP Human - LCL

Get tips on using EpiTect ChIP qPCR Assays to perform ChIP Mouse - NIH3T3

Get tips on using EpiTect ChIP OneDay Kit to perform ChIP Human - HUH-7

Get tips on using EpiTect ChIP OneDay Kit to perform ChIP Human - MIA PaCa-2

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.

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 TruSeq ChIP Library Preparation Kit to perform ChIP Mouse - Ovary

Get tips on using Chromatin Immunoprecipitation (ChIP) Assay Kit to perform ChIP Mouse - Ovary

Get tips on using Pierce™ Magnetic ChIP Kit to perform ChIP Mouse - C2C12

Get tips on using Chromatin Immunoprecipitation (ChIP) Assay Kit to perform ChIP Mouse - C2C12