siRNA / miRNA gene silencing Human Capan-1

Get tips on using Endothelin 1 ELISA Kit (ab133030) to perform ELISA Rat - Endothelin 1

Get tips on using Beclin-1 Antibody to perform Autophagy assay cell type - NRK-49F

Get tips on using Beclin-1 Antibody to perform Autophagy assay cell type - BRL-3A

Get tips on using pLKO.1 puro to perform CRISPR Mouse - Activation RAW 264.7 Dmd

Get tips on using pLKO.1 puro to perform CRISPR Mouse - Deletion 3T3-L1 Epac1

Get tips on using NucleoSpin® miRNA to perform RNA isolation / purification Cells - primary canine aortic endothelial cells

Get tips on using pGEX-4T-1 to perform Protein Expression Prokaryotic cells - E. coli LsrK

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.

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 Beclin-1 (D40C5) Rabbit mAb to perform Autophagy assay cell type - THP 1