siRNA / miRNA gene silencing Human Primary Endometrial Stromal Cells hsa-miR-542-3p

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.

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

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

Get tips on using Human TGF-beta 1 Quantikine ELISA Kit to perform ELISA Human - TGF-beta 1

Get tips on using Human IL-1 beta ELISA Kit (ab100562) to perform ELISA Human - IL-1 beta

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 PE anti-human CD114 (G-CSFR) Antibody to perform Flow cytometry Anti-bodies Human - CD114

Get tips on using PE Mouse Anti-Human CD26 Clone L272 to perform Flow cytometry Anti-bodies Human - CD26

Get tips on using PE Mouse Anti-Human CD30 Clone BerH8 to perform Flow cytometry Anti-bodies Human - CD30