CRISPR Mouse Deletion RMA cells

Get tips on using Dulbecco’s Modified Eagle’s Medium/Ham’s Nutrient Mixture F12 to perform 3D Cell Culture Media PLC/PRF/5 cells-spheres

Get tips on using Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham to perform Stem cell culture media NCH421K cells primary glioma

Get tips on using The Premo Autophagy Tandem Sensor RFP-GFP-LC3B Kit to perform Autophagy assay cell type - Human osteosarcoma cancer 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.

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 mirVana® miRNA mimic to perform siRNA / miRNA gene silencing Human - Primary Endometrial Stromal Cells hsa-miR-542-3p

Get tips on using pwPICZalpha-DT390-bi-pIL-2-Gly to perform Protein Expression Eukaryotic cells - P. pastoris Porcine IL-2 fusion toxins

Get tips on using pFastBac-GP67-H6HA1-His-RhPV-IRES-EGFP to perform Protein Expression Eukaryotic cells - S. frugiperda HA1 of H6N1 AIV

Get tips on using GeneChip® Human Genome U133 Plus 2.0 Array to perform Microarray Gene expression arrays - Human endometrial stromal cells Biotin

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.