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G418 Disulfate

nr kat.: G001-25G
Opakowanie: 25 g
Cena brutto: 2 249,67 zł 2249.67
Cena netto: 1 829,00 zł
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Producent: TOKU-E
nr kat.: G001-25G

Opis

Produkt przeznaczony wyłącznie do badań. Nie nadaje się do spożycia. Więcej informacji na stronie producenta www.toku-e.com


G418 disulfate, also known as G418 sulfate, is routinely used as a selection antibiotic in cell culture gene selection applications. G418 disulfate is an aminoglycoside antibiotic isolated from Micromonospora rhodorangea and is closely related to the gentamicins; most notably, gentamicin B1.

 TOKU-E also offers:

  • G418 disulfate solution (G021)
  • G418 disulfate, EvoPure (G030) (≥99.0% pure)
  • G418 disulfate, (low endotoxin) (G048)


Suasnavas et al. used G418 disulfate from TOKU-E to select for resistant porcine TE and FF cells. Read more here: "Characterization and Potential Utility of Porcine Trophoblast-Derived Stem-Like Cells."

Wang, et al. used G418 disulfate from TOKU-E in RPMI media to select RFP expressing TC-2 cells. Read more here: "3D Porous Chitosan–Alginate Scaffolds as an In Vitro Model for Evaluating Nanoparticle-Mediated Tumor Targeting and Gene Delivery to Prostate Cancer."

Mechanism of Action
G418 disulfate, and other aminoglycosides, including kanamycin and neomycin, prevent protein synthesis by blocking the elongation step in prokaryotic and eukaryotic ribosomes.

Mechanism of resistance:
Resistance to G418 sulfate is conferred by the neo gene (neomycin resistant gene) from either Tn5 or Tn601 (903) transposons. Cells successfully transfected with resistance plasmids containing the neo resistance gene can express aminoglycoside 3'-phosphotransferase (APT 3' I or APT 3' II) which covalently modifies G418 to 3-phosphoric G418. 3-phosphoric G418 has negligible potency and has low-affinity for prokaryotic or eukaryotic ribosomes.

Spectrum
G418 disulfate is toxic to susceptible prokaryotic and eukaryotic cells including fungi (yeasts and molds), bacteria, mammalian and plant cells.

Microbiology Applications
G418 Disulfate can be used as a selection agent for G418 resistant bacteria or fungi after transformation.

Eukaryotic Cell Culture Applications
G418 sulfate is routinely used as a selection agent in cell culture after transfection of eukaryotic cells. G418 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 sulfate and other aminoglycoside antibiotics.

Optimal working concentration ranges of G418 sulfate are as follows:

  • Mammalian cell lines – 200 mg/L – 1000 mg/L
  • Bacteria and algae - ≤5 mg/L or less for 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.

Note: The optimal working concentration of G418 sulfate to select for resistant mammalian clones depends on the cell lines used, the quality of G418 sulfate, media, growth conditions, cell density, cell metabolic rate, cell cycle phase, and the plasmid carrying the neo resistance gene. A kill curve should therefore be performed to determine the optimal working concentration for every experimental system and for every lot of G418 sulfate. A kill curve can be performed using the following concentrations of G418 sulfate:
        5 mg/L - 1400 mg/L for mammalian cells
        0.1 mg/L - 50 mg/L for bacteria (E. Coli ) and algae.

High quality G418 sulfate ideally has high potency against sensitive cells (low ED50 concentration) and low potency against transfected resistant cells (high ED50 concentration). Impurities and certain bio active substances such as endotoxins present in G418 sulfate can be toxic to resistant cells and can cause death at relatively low concentrations.

TOKU-E G418 stock solutions are prepared at 50mg/mL for quick and easy dilution factor calculations

For more information on relevant cell lines, culture medium, and working concentrations, please visit the TOKU-E Cell-culture Database.

Suasnavas et al. used G418 disulfate from TOKU-E to select for resistant porcine TE and FF cells. Read more here: "Characterization and Potential Utility of Porcine Trophoblast-Derived Stem-Like Cells."

References
Aragão F.J.L. and Brasileiro A.C.M., Positive, negative and marker-free strategies for transgenic plant selection. Braz. J. Plant Physiol., 14(1):1-10, 2002
Davis, Bernard D. "Mechanism of Bactericidal Action of Aminoglycosides."Microbiological Reviews 51.3 (1987): 341-50.
Lin-Cereghino, Joan, et. al.. "Direct Selection of Pichia pastoris Expression Strains Using New G418 Resistance Vectors." Yeast 25 (2008): 293-99. Interscience.wiley.com. Web. 25 Apr. 2013.
Shin, Young-Mi. "Selection of NptII Transgenic Sweetpotato Plants Using G418 and Paromomycin." Journal of Plant Biology 50.2 (2007): 206-12. Researchgate.net. Web. 25 Apr. 2013.

Dane techniczne

Opakowanie 25 g

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