crispr-mouse-deletion-raw-264-7-dcstamp

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

Get tips on using Cell Death Detection ELISA to perform TUNEL assay cell type - Rat pulmonary arterial smooth muscle cells

Get tips on using Ras (D2C1) Rabbit mAb #8955 to perform Western blotting Ras

Get tips on using TdT In Situ Apoptosis Detection Kit - Fluorescein to perform TUNEL assay cell type - Rat fibroblast-like synoviocytes

Get tips on using ROS-ID® Total ROS/Superoxide detection kit to perform ROS assay cell type - H9c2 rat cardiomyocytes

Get tips on using DCFDA - Cellular Reactive Oxygen Species Detection Assay Kit to perform ROS assay cell type - H9c2 rat cardiomyocytes

Get tips on using In Situ Cell Death Detection Kit, TMR red to perform TUNEL assay cell type - Rabbit synovial fibroblasts

Get tips on using DCFDA - Cellular Reactive Oxygen Species Detection Assay Kit to perform ROS assay cell type - rat kidney and pancreas tissue

Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

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.