Protein Expression Eukaryotic cells A. thaliana

Get tips on using CelLytic™ M to perform Protein isolation Mammalian cells - SK-N-BE(2)-C

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

Get tips on using IGEPAL® CA-630 to perform Protein isolation Mammalian cells - Human lung fibroblasts

Get tips on using CelLytic™ NuCLEAR™ Extraction Kit to perform Protein isolation Mammalian cells - HLE-B3

Get tips on using CelLytic™ NuCLEAR™ Extraction Kit to perform Protein isolation Mammalian cells - MLS-1765

Get tips on using CelLytic™ NuCLEAR™ Extraction Kit to perform Protein isolation Mammalian cells - KC02-44D

Get tips on using CelLytic™ NuCLEAR™ Extraction Kit to perform Protein isolation Mammalian cells - BHK-21

Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Tissue - Human aortic endothelial cells

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.

Isolating DNA from tissues and paraffin-embedded tissue samples can be challenging as double-stranded DNA is physically fragile and highly susceptible to exo- and endonucleases. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in the presence of DNAse inhibitors. Further, extracting DNA from the nucleus need specific methods by combining physical, mechanical and chemical lysis approaches,