siRNA / miRNA gene silencing Mouse CT26

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 - NIH-3T3

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.

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.

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.