Protein expression and purification Mammalian cells HEK 293

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary porcine primary airway epithelial cell

Get tips on using X-VIVOTM 10 Serum-free Hematopoietic Cell Medium to perform Mammalian cell culture media NK-92

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform siRNA / miRNA gene silencing Human - Primary Endometrial Stromal Cells IGFBP1 (Insuline-like growth factor binding protein-1) Lipid

Get tips on using Senescence Cells Histochemical Staining Kit to perform Reporter gene assay β-galactosidase substrates - adipose stem cells

Get tips on using Dulbecco’s Modified Eagle’s Medium (DMEM) to perform Mammalian cell culture media HepG2

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.

Get tips on using Blood & Cell Culture DNA Midi Kit (25) to perform DNA isolation / purification Cells - Immortalized cell lines Human Neuroblastoma Cell Lines

Get tips on using DeadEnd™ Colorimetric TUNEL System to perform TUNEL assay cell type - HNSCC Detroit 562 human head and neck tumor cells

Get tips on using Prestained Protien ladder to perform Protein Ladder Prestained

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.