Description
Nycodenz® is the ideal density separation medium for the fractionation of viruses, diverse subcellular organelles, membranes, macromolecules, polysaccharides, nucleoproteins and cells. Its low osmolality and non-toxicity recommend it for separating living cells. Its low viscosity and non-ionic nature have proven useful in the isolation and purification of viruses and bacteria.
Nycodenz®, the trademark name for iohexol whose systematic name is 5-(N-2, 3-dihydroxypropylacetamido)-2, 4, 6-tri-iodo-N, N'-bis (2,3-dihydroxypropyl) isophthalamide, was originally developed as an X-ray contrast medium and has therefore been subjected to rigorous clinical testing. It is a non-ionic, tri-iodinated derivative of benzoic acid with three aliphatic hydrophilic side chains. Its high solubility, underlying its ability to generate high density solutions, derives from the hydrophilic groups linked to its triiodobenzene ring. The dihydroxypropylacetamido side chain is responsible for its very low toxicity compared to metrizamide. The iodinated aromatic nucleus absorbs strongly in the ultraviolet region of the spectrum with an absorption maximum of 244 nm.
Applications
- Mammalian and non-mammalian cell isolation
- Virus isolation
- Subcellular organelle isolation
- Subcellular membrane isolation
- Protein and protein complex isolation
- Ribonucleoprotein isolation
- Macromolecule isolation
Features and properties
- Molecular weight: 821 Da
- Supplied as a powder
- Non-ionic
- Non-toxic to cells
- Metabolically inert
- Resistant to bacterial degradation
- Autoclavable
- Forms true solutions, permitting dialysis, ultrafiltration, or gel filtration for easy removal from cells after fractionation
- Amenable to density gradient generation by:
- Diffusion of a discontinuous gradient — simple, sub-45-minute preparation time
- Tilted tube rotation (Gradient Master™)
- Two-chamber gradient maker
- Freezing/thawing of a uniform density solution
- Centrifugation (auto-generated gradient)
- Solution density can be determined from measurement of refractive index or absorbance
- Non-particulate nature permits measurement of cellular distribution across gradient fractions by hemocytometer, electronic particle counter or by spectrophotometric light scattering
- Compatible with the orcinol and diphenylamine reactions and with very sensitive dye binding and fluorometric assays for estimation of nucleic acid and protein concentrations
- Compatible with most subcellular component marker enzyme assays
- Compatible with commercial scintillants
- Compatible with the phenol/H2SO4 assay for detection of mono and polysaccharides
- Compatible with commercial scintillants
- Compatible with the phenol/H2SO4 assay for detection of mono and polysaccharides
Stability and storage
Nycodenz® in solid form is stable for a period of 5 years when stored at RT and protected from light. In solution it is stable for 5 years provided it is kept sterile and protected from light. Although, prolonged exposure to direct sunlight leads to release of iodine from the molecule, this effect is negligible when working with solutions on a day-to-day basis.
Optiprep™/ Nycodenz® Application sheets
