更新時間：2022-01-29
G418 also known as G-418, G418 sulfate or Geneticin is an aminoglycoside antibiotic produced by Micromonospora rhodorangea.G418 blocks polypeptide synthesis by inhibiting the elongation step in both…
G418 also known as G-418, G418 sulfate or Geneticin is an aminoglycoside antibiotic produced by Micromonospora rhodorangea.
G418 blocks polypeptide synthesis by inhibiting the elongation step in both prokaryotic and eukaryotic cells.
Resistance to G418 is conferred by the Neomycin resistance gene (neo) from Tn5 encoding an aminoglycoside 3'-phosphotransferase, APH 3' II.
Selection in mammalian cells is usually achieved in 3 to 7 days with concentrations ranging from 400 to 1000 µg/ml.
Cells that are dividing are affected more rapidly than non-dividing cells.
Product concentration: 100 mg/ml
CAS number: 108321-42-2
Quality Control: Each lot is thoroughly tested to ensure the absence of lot-to-lot variation.
Purity: ≥ 90% (HPLC)
Endotoxin level: < 0.5 EU/mg
Physicochemical characterization: pH, appearance
Cell-culture tested: potency validated in G418-sensitive and G418-resistant mammalian cell lines
Non-cytotoxicity of trace contaminants: absence of long-term effects confirmed in G418-resistant cells
Formula: C20H40N4O10. 2H2SO4
Molecular weight: 692.7
Structure:
G418 is supplied as a sterile filtered solution at 100 mg/ml in HEPES buffer.
This product is available in three pack sizes:
ant-gn-1: 10 x 1 ml (1 g)
ant-gn-2: 20 x 1 ml (2 g)
ant-gn-5: 1 x 50 ml (5 g)
G418 is shipped at room temperature.
Upon receipt it should be stored at 4°C.
G418 is normally used at a concentration of 400 μg/ml. However, the optimal concentration needs to be determined for your cells.
Suggested concentrations of G418 for selection in some examples of mammalian cells are listed below:
Cell line | Medium | G418 conc | References |
|---|---|---|---|
| B16 (Mouse melanocytes) | RPMI | 400-1000 μg/ml | 1, 2 |
CHO (Chinese hamster ovarian cells) | Ham’s | 400-800 μg/ml | 3 |
| HeLa (Human uterine cells) | DMEM | 200-400 μg/ml | 4, 5 |
| HEK293 (Human embryonic kidney cells) | DMEM | 200-500 μg/ml | 6, 7 |
| THP-1 (Human monocytes) | RPMI | 250 μg/ml | 8, 9 |
References:
1. Arikawa K. et al., 2003. Ligand-dependent inhibition of B16 melanoma cell migration and invasion via endogenous S1P2 G protein-coupled receptor. Requirement of inhibition of cellular RAC activity. J Biol Chem. 278:32841-51.
2. DiLillo DJ. et al., 2010. B cells are required for optimal CD4+ and CD8+ T cell tumor immunity: therapeutic B cell depletion enhances B16 melanoma growth in mice. J Immunol. 184:4006-16.
3. Figueroa K. et al., 2009. Selectivity of agonists for the active state of M1 to M4 muscarinic receptor subtypes. J Pharmacol Exp Ther. 28:331-42.
4. Kwon JA. et al., 2005. Biological function of the vaccinia virus Z-DNA-binding protein E3L: gene transactivation and anti-apoptotic activity in HeLa cells.
5. Hermoso M. et al., 2004. Cell volume regulation in response to hypotonicity is impaired in HeLa cells expressing a protein kinase Calpha mutant lacking kinase activity. J Biol Chem. 279:17681-9.
6. Hennen S. et al., 2013. Decoding signaling and function of the orphan G protein-coupled receptor GPR17 with a small-molecule agonist. Sci. Signal., 6:ra93.
7. Chen Q. et al., 2003. Expression of Drosophila trehalose-phosphate synthase in HEK-293 cells increases hypoxia tolerance. J Biol Chem. 278:49113-8.
8. Tukhvatulin A. et al., 2013. Combined stimulation of Toll-like receptor 5 and NOD1 strongly potentiates activity of NF-κB, resulting in enhanced innate immune reactions and resistance to Salmonella enterica serovar Typhimurium infection. Infect Immun. 81:3855-64.
9. Maue AC. et al., 2013. The polysaccharide capsule of Campylobacter jejuni modulates the host immune response. Infect Immun. 81:665-72.
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