G418 Disulfate, EvoPure®

- Overview
  - G418 Disulfate, EvoPure® is a highly pure (≥99.0%) version of G418 Disulfate. It has been purified to remove the impurities commonly present in G418 Disulfate. This aminoglycoside antibiotic, originally isolated from Micromonospora rhodorangea, is routinely used for gene selection in cell culture.
    
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    Background
    
    G418 Disulfate, and other aminoglycosides, including Kanamycin and Neomycin, prevent protein synthesis by blocking the elongation step in prokaryotic and eukaryotic ribosomes.
- Properties
  - CAS Number
    
    108321-42-2
    
    Molecular Formula
    
    C20H40N4O10 · xH2SO4 (lot specific)
    
    Molecular Weight
    
    496.55 g/mol (Free base)
    
    Other Properties
    
    Source: Biosynthetic: produced by Micromonospora rhodorangea.
    PH: 4.6-6.0
    Absorbance: 1mg/ml (water): 280nm <0.015 570nm <0.01 100mg/ml (water): 570nm <0.01 1.74g/25 ml (water): 280nm <0.7
    Boiling Point: 1012.1 °C
    
    Storage
    
    ≤30 °C
    
    * For research use only
- Applications
  - Application Description
    
    Spectrum: G418 Disulfate is toxic to susceptible bacteria and fungi
    
    Microbiology Applications: G418 Disulfate is used as a gene selection agent during transfection of eukaryotic cells.
    
    Eukaryotic Cell Culture Applications: G418 Disulfate is routinely used as a selection agent in cell culture after transfection of eukaryotic cells. Resistant cells express the neo gene which produces aminoglycoside 3'-phosphotransferase (APT 3' I or APT 3' II), a protein that confers resistance to G418 Disulfate and other aminoglycoside antibiotics.
    Optimal working concentrations:
    
    Mammalian cell lines: 200 mg/L – 1000 mg/L
    Bacteria and algae: ≤5 mg/L
    
    The optimal working concentration of G418 Sulfate to select for resistant clones depends on the cell line, reagent quality, reagent lot, media, growth conditions, cell density, cell metabolic rate, cell cycle phase, and plasmid quality. A kill curve should be performed to determine the optimal concentration for each experimental system.
    Use the following guide to determine the concentration to use to generate a kill curve:
    
    5 mg/L - 1400 mg/L (mammalian cells)
    0.1 mg/L - 50 mg/L (bacteria and algae)
    
    A working concentration of 200 mg/L is usually sufficient after resistant mammalian clones are selected and can be used for maintenance until stable resistant clones are selected.
    The Selectivity Factor is a quantifiable measure of how efficient an antibiotic is during the process of gene selection. Our technical team tested the selectivity factor of G418 for BHK-21 cells and HeLa cells. We found that G418 is an ideal selection antibiotic for transfected BHK-21 cells but not optimal for HeLa cells. The method uses a modified MTT assay, which can be used to numerically determine the antibiotic efficiency (Delrue I et al, 2018). .
    For more information on relevant cell lines, culture medium, and working concentrations, please visit the TOKU-E Cell-culture Database.