siRNA / miRNA gene silencing Human ACC-MESO1

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 Human/Mouse/Rat/Canine ALCAM/CD166 Antibody to perform Immunohistochemistry Mouse - CD166 / ALCAM

Get tips on using Human ICAM-1/CD54 Antibody to perform Western blotting ICAM-1

As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.

As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.

Get tips on using RosetteSep™ Human T Cell Enrichment Cocktail to perform Cell Isolation Human T cells

Get tips on using MagniSort™ Human T cell Enrichment Kit to perform Cell Isolation Human T cells

Get tips on using EasySep™ Human T Cell Enrichment Kit to perform Cell Isolation Human T cells

Get tips on using EasySep™ Human T Cell Isolation Kit to perform Cell Isolation Human T cells

Get tips on using Human TGF beta 1 ELISA Kit (ab100647) to perform ELISA Human - TGF-beta 1