Protein Expression Eukaryotic cells S. pastorianus

Get tips on using Cell Lysis Buffer (10X) to perform Protein isolation Mammalian cells - THP-1

Get tips on using Cell Lysis Buffer (10X) to perform Protein isolation Mammalian cells - KC02-44D

Get tips on using NP40 Cell Lysis Buffer to perform Protein isolation Mammalian cells - BHK-21

Get tips on using RIPA Lysis and Extraction Buffer to perform Protein isolation Mammalian cells - HepG2

Get tips on using RIPA Lysis and Extraction Buffer to perform Protein isolation Mammalian cells - HEK293T

Get tips on using QuantiPro™ BCA Assay Kit to perform Protein quantification Mammalian cells - L929

Get tips on using QuantiPro™ BCA Assay Kit to perform Protein quantification Mammalian cells - RAW264.7

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Mammalian cells - HUVEC

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Mammalian cells - HeLa

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.