siRNA / miRNA gene silencing Rat Brain endothelial cells HIF-1α

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 Anti-LAMP1 antibody [1D4B] (ab25245) to perform Autophagy assay cell type - RAW 264.7

Get tips on using Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate to perform Flowcytometry Secondary Antibody - Goat Rabbit Alexa Fluor 488

Get tips on using CYTO-ID® Autophagy detection kit to perform Autophagy assay cell type - Ramos

Get tips on using APO-BRDU™ Kit (RUO) to perform TUNEL assay cell type - Rabbit synovial fibroblasts

Get tips on using Hin1II (NlaIII) (5 U/µL) to perform Restriction Enzymes Hin1II / NlaIII

Get tips on using Annexin V-FITC Apoptosis Detection Kit to perform Apoptosis assay cell type - RAW 264.7

Get tips on using Ni-NTA HisSorb Strips (24) to perform Protein tag Detection of His-tagged proteins

Get tips on using Anti-phospho-Histone H2A.X (Ser139) Antibody to perform TissueFAxs phospho-Histone H2A.X (Ser139) - Mouse Human

Get tips on using Anti-phospho-Histone H2A.X (Ser139) Antibody to perform Immunofluorscence  phospho-Histone H2A.X (Ser139) - Mouse Human