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.
Get tips on using PE Mouse Anti-Human CD31 Clone L133.1 to perform Flow cytometry Anti-bodies Human - CD31/PECAM-1
Get tips on using Monoclonal Mouse Anti-Human Actin (Smooth Muscle) (Concentrate) Clone 1A4 to perform Immunohistochemistry Mouse - SMA
Get tips on using Human/Mouse GFR alpha-2/GDNF R alpha-2 Antibody to perform Immunohistochemistry Mouse - Gfrα2
Get tips on using Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor® 568 conjugate to perform Flowcytometry Secondary Antibody - Goat Mouse Alexa Fluor 568
RNA-Seq is a method to sequence RNA by applying Next Generation Sequencing (NGS). The quality of RNA is critical for the success of RNA-Seq. The integrity of RNA is measured by the RNA integrity number (RIN). RIN is computed from RNA electrophoresis and electropherogram profiles (the peak area of the 28S rRNA should be approximately twice the peak area of the 18S rRNA). If you get the RIN value lower than 7, the possibility of getting the low quality of RNA-seq data is high. To get a high quality RNA, it is better to work with fresh samples or snap-freeze the tissues in liquid nitrogen as quickly as possible and store them at -80°C until further use. Make sure designated areas and all your filter tips, microfuge tubes, plastic, and glassware are RNase-free.
Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase
Get tips on using Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA Kit to perform ELISA Mouse - TGF-beta 1
Get tips on using Mouse CRP / C Reactive Protein / PTX1 PicoKine™ ELISA Kit to perform ELISA Mouse - C-Reactive Protein/CRP
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