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 anti-alpha-Smooth Muscle Actin mouse monoclonal, ASM-1 to perform Immunohistochemistry Alpha smooth muscle Actin - Mouse -NA- -NA-
Get tips on using Phospho-ULK1 (Ser757) Antibody to perform Autophagy assay cell type - Human primary MSCs
Get tips on using LC3B Antibody Kit for Autophagy to perform Autophagy assay cell type - Human melanocytes
Get tips on using ROS-ID® Total ROS/Superoxide detection kit to perform ROS assay cell type - A549 human adenocarcinomic human alveolar basal epithelial cells
Get tips on using Fluorometric Intracellular Ros Kit to perform ROS assay cell type - SH-SY5Y human neuroblastoma
Get tips on using Cell Death Detection ELISA to perform Apoptosis assay cell type - Human endometrial stromal cells
Get tips on using In situ apoptosis detection to perform Apoptosis assay cell type - Human endometrial stromal cells
Get tips on using IncuCyte® Cell Migration Kit to perform Wound healing assay cell type - human MEWO
Get tips on using IncuCyte® Cell Migration Kit to perform Wound healing assay cell type - human A375
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment