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Get tips on using Amino Allyl MessageAmp™ II aRNA Amplification Kit to perform Microarray RNA amplification & Labeling - Human brain tissue Cyanine 3

Get tips on using Amino Allyl MessageAmp™ II aRNA Amplification Kit to perform Microarray RNA amplification & Labeling - Fish fundulus heteroclitus Cyanine-3 / Cyanine-5

Get tips on using Amino Allyl MessageAmp™ II aRNA Amplification Kit to perform RNA amplification & labeling Fish - Total RNA, Fundulus heteroclitus Cyanine 3 & 5

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.

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.

Get tips on using Anti-REG1 alpha antibody (ab231092) to perform Immunohistochemistry Mouse - Reg1

Get tips on using Anti-Choline Acetyltransferase Antibody AB5042 to perform Immunohistochemistry Mouse - ChAT

Get tips on using Anti-activated Notch1 antibody (ab8925) to perform Western blotting Notch1

Get tips on using Anti-LC3B antibody (ab48394) to perform Autophagy assay cell type - Cholangiocarcinoma