siRNA / miRNA gene silencing Mouse NIH-3T3

Get tips on using PerCP-Cy™5.5 Mouse Anti-Human CD19 to perform Flow cytometry Anti-bodies Human - CD19

Get tips on using APC-Cy™7 Mouse Anti-Human CD19 to perform Flow cytometry Anti-bodies Human - CD19

Get tips on using APC-Cy™7 Mouse Anti-Human CD3 to perform Flow cytometry Anti-bodies Human - CD3

Get tips on using Anti-LGR5 mouse mAb, clone OTI2A2, PE conjugated to perform Flow cytometry Anti-bodies Human - LGR5

Get tips on using Alexa Fluor® 647 Mouse Anti-Human CD24 to perform Flow cytometry Anti-bodies Human - CD24

Get tips on using Purified Mouse Anti-Beclin Clone 20/Beclin (RUO) to perform Autophagy assay cell type - THP 1

A standard angiogenic assay involves the autonomous endothelial cell response of self-organization into microvessels, also known as tubes when seeded on a basement membrane matrix in the presence of the appropriate growth factors. However, the component of basement membrane matrix may also affect the tube formation by endothelial cells. Hence it is important to use a standard angiogenesis assay kit or use the same membrane matrix with known composition to standardize the assay conditions.

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 Purified Mouse Anti-SV40 Large T Antigen Clone PAb 101 (RUO) to perform Immunohistochemistry Mouse - SV40

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.