DNA transfection Mammalian cells Immortalized cell lines

Get tips on using Minimum Essential Medium Eagle to perform Mammalian cell culture media BAOSMC

Get tips on using MEM (Eagle) Liquid Medium to perform Mammalian cell culture media MDCK

Get tips on using Minimum Essential Media (MEM) to perform Mammalian cell culture media MDCK

Get tips on using Minimum Essential Media (MEM) to perform Mammalian cell culture media Vero

Get tips on using Minimum Essential Medium Eagle to perform Mammalian cell culture media Vero

Get tips on using MultiTox-Fluor Multiplex Cytotoxicity Assay to perform Live / Dead assay mammalian cells - RAW 264.7

Get tips on using AllPrep DNA/RNA Micro Kit to perform RNA isolation / purification Tissue - Human Blood / Serum / Plasma / Buffy coat

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.

Cell Invasion or Cell Migration assays are technically challenging to set up as the gradient between the two compartments equilibrates in time during the assay. It is also problematic to view cells and for cells to migrate through a non-physiologic polycarbonate or polypropylene filter. Care must be taken while loading the well with cells to form a single cell suspension. Precaution must be taken while trypsinization (under-trypsinization can lead to cell clumping while over-trypsinization could strip off adhesion molecules necessary for migration). This leads to difficulty in getting significant results, when only small numbers of cells cross the filter or when the distribution and/or staining of the cells is uneven