| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
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| Other Sizes |
| Targets |
Aminoglycoside
G-418 targets the ribosome, specifically inhibiting the elongation step of protein synthesis in both prokaryotic and eukaryotic cells. As an aminoglycoside antibiotic, it binds to the 30S ribosomal subunit in prokaryotes and the 40S subunit in eukaryotes, causing misreading of the genetic code and inhibiting polypeptide chain elongation. The neomycin resistance gene (neo) encodes an aminoglycoside phosphotransferase that detoxifies G-418, allowing for selection of transfected cells. |
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| ln Vitro |
G-418 (500 μg/mL, 12 days) is employed in drug-resistant LNCaP cell screening [7].
In vitro, G-418 (500 μg/mL; 12 days) is employed in drug-resistant LNCaP cell screening. It inhibits a variety of prokaryotic and eukaryotic organisms at concentrations ranging from 1 to 300 μg/mL. In Acanthamoeba, G-418 activates programmed cell death, causing shape changes, cell rounding, increased intracellular calcium (from 24 nM to 60 nM after 6 hours), mitochondrial dysfunction, cytochrome c release, and nuclear vesiculation. The antibiotic's broad-spectrum activity makes it valuable for selection and screening applications. |
| ln Vivo |
Geneticin G418 (G-418 disulfate) (40 and 80 mg/kg) for three consecutive days are sufficient to eliminate all nontransfected T. brucei brucei parasites from infected mice[3].
In vivo, G-418 (as G-418 disulfate) at 40 and 80 mg/kg for three consecutive days is sufficient to eliminate all nontransfected T. brucei brucei parasites from infected mice. In mouse protection tests, antibiotic G-418 is approximately half as potent as gentamicin. The compound has also been used to study nonsense suppression in Mc4r(X16) knock-in mouse models. These in vivo applications demonstrate the utility of G-418 for selection in transgenic animal models and antiparasitic research. |
| Enzyme Assay |
The in vitro antimicrobial susceptibility assay for G-418 involves testing the compound against various prokaryotic and eukaryotic organisms using standard broth microdilution methods. Minimum inhibitory concentrations (MICs) are determined after 16-24 hours of incubation at appropriate temperatures. For mammalian cell selection, cells are cultured in medium containing G-418 at concentrations typically ranging from 200-1000 μg/mL, depending on the cell line and resistance gene. Selection is maintained for 7-14 days until resistant colonies appear. Cytotoxicity is assessed using standard cell viability assays.
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| Cell Assay |
Acanthamoeba is a widely distributed opportunistic parasite which causes a vision-threatening keratitis and a life-threatening encephalitis. The cyst stage of this amoeba is especially resistant to currently used therapeutics and so alternative agents are urgently required. Growing evidence supports the existence of a programmed cell death system (PCD) in Acanthamoeba and while some features are shared by higher eukaryote cells, others differ. It is hoped that by understanding these differences we can exploit them as targets for novel drug intervention to activate PCD pathways in the amoebae but not the invaded human tissue. Here, we use the aminoglycoside G418 to activate PCD in Acanthamoeba. This drug caused a shape change in the treated amoebae. Cells rounded up and contracted, and after 6 h fragments of cells resembling the 'apoptotic bodies' of vertebrate cells were observed. G418 causes an increase in intracellular calcium from a resting level of 24 nM to 60 nM after 6 h of treatment. Mitochondrial function as assayed by the ΔΨm reporting dye JC-1 and CTC a redox dye becomes inhibited during treatment and we have found that cytochrome c is released from the mitochondria. Cells stained with Hoechst showed first an alteration in chromatin structure and then a vesiculation of the nucleus with G418 treatment, although we found no obvious breakdown in genomic DNA in the early stages of PCD [5].
Cellular assays for G-418 are conducted in various mammalian cell lines for selection of transfected cells. Cells are transfected with a vector containing the neomycin resistance gene and then cultured in medium containing G-418 at concentrations of 200-1000 μg/mL. Selection is maintained for 7-14 days, and resistant colonies are counted or expanded. In Acanthamoeba studies, G-418 is used to induce programmed cell death, and effects are assessed by microscopy, calcium imaging, mitochondrial membrane potential assays (JC-1), and cytochrome c release assays. |
| Animal Protocol |
Mouse RML and 22L prion inocula were obtained from RML- or 22L-infected CAD5 cells (34). The cells were grown to confluency, scraped in 1 ml PBS per 10 cm dish, and then homogenized using a Minilys bead homogenizer and CK14 homogenization tubes (Bertin). Benzonase (50 units/ml; EMD Millipore #70746-4) was added into the scraped culture, and then the culture was homogenized for three cycles of 30 s at maximum speed, with 5 min of incubation on ice between each cycle. Cell homogenates were then stored at −80 °C. Syrian hamster 263K prion inoculum was prepared in a similar way from 263K-infected CAD5-PrP−/− cells stably expressing HaPrP.[6]
In vivo studies for G-418 are conducted in mouse models. For parasite elimination studies, mice infected with T. brucei brucei are treated with G-418 disulfate at 40 or 80 mg/kg for three consecutive days. Efficacy is assessed by measuring parasite clearance from infected animals. For nonsense suppression studies, Mc4r(X16) knock-in mice are used to evaluate restoration of Mc4r function. For prion studies, mouse RML and 22L prion inocula are prepared from infected CAD5 cells. |
| ADME/Pharmacokinetics |
G-418 is typically administered as the disulfate salt for in vivo studies. In mouse models, G-418 disulfate is administered at 40 or 80 mg/kg for three consecutive days. The compound has a molecular weight of 496.55 g/mol and a molecular formula of C₂₀H₄₀N₄O₁₀. Storage conditions: powder at -20°C or 4°C. The compound is soluble in water and common buffers. Detailed PK parameters such as half-life and bioavailability are not extensively reported in the available literature for this research compound.
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| Toxicity/Toxicokinetics |
In mouse protection tests, G-418's acute toxicity is one-half to one-third that of gentamicin. At the studied doses of 40 and 80 mg/kg for three consecutive days, the compound was effective in eliminating parasites. As an aminoglycoside antibiotic, potential toxicities include nephrotoxicity and ototoxicity, similar to other aminoglycosides. As with all research compounds, G-418 is intended for research use only and not for human therapeutic applications. Appropriate safety precautions should be taken when handling this compound.
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| References |
[1]. An antibiotic selection marker for nematode transgenesis. Nat Methods. 2010;7(9):721-723.
[2]. A novel G418 conjugate results in targeted selection of genetically protected hepatocytes without bystander toxicity. Bioconjug Chem. 2007;18(6):1965-1971. [3]. Disialyl GD2 ganglioside suppresses ICAM-1-mediated invasiveness in human breast cancer MDA-MB231 cells. Int J Biol Sci. 2017;13(3):265-275. Published 2017 Feb 12. [4]. A new selective agent for eukaryotic cloning vectors. Am J Trop Med Hyg. 1980;29(5 Suppl):1089-1092. [5]. G418 induces programmed cell death in Acanthamoeba through the elevation of intracellular calcium and cytochrome c translocation. Parasitol Res. 2019 Feb;118(2):641-651. [6]. J Biol Chem. 2021 Sep;297(3):101073. doi: 10.1016/j.jbc.2021.101073. Epub 2021 Aug 12. [7]. S Bennett, et al. Reliable method of isolating transfected clones from the LNCaP human prostatic cell line. Biotechniques. 1997 Jul;23(1):66, 68, 70. |
| Additional Infomation |
Geneticin (also known as G418) is an aminoglycoside antibiotic with a structure similar to gentamicin B1, produced by Micromonospora rhodorangea. Geneticin blocks polypeptide synthesis by inhibiting the elongation step of polypeptide chains in both prokaryotic and eukaryotic cells, and is commonly used in laboratory research to screen genetically engineered cells. Resistance to Geneticin is conferred by the neo gene on the Tn5 transporter, which encodes the aminoglycoside 3'-phosphotransferase APH 3' II.
G-418 (Geneticin) is an aminoglycoside antibiotic produced by Micromonospora rhodorangea that inhibits protein synthesis by blocking the elongation step. It is toxic to both eukaryotic and prokaryotic cells and is widely used as a selection agent for mammalian cell lines expressing the neomycin resistance gene. In vitro, G-418 is used at 500 μg/mL for drug-resistant cell screening. In vivo, G-418 disulfate at 40-80 mg/kg eliminates nontransfected parasites from infected mice. Its molecular formula is C₂₀H₄₀N₄O₁₀ with a molecular weight of 496.55 g/mol. G-418 is an essential tool for molecular biology and genetic research. |
| Molecular Formula |
C20H40N4O10
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|---|---|
| Exact Mass |
496.274
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| CAS # |
49863-47-0
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| Related CAS # |
G-418 disulfate;108321-42-2
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| PubChem CID |
123865
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.47 g/cm3
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| Boiling Point |
760.3ºC at 760 mmHg
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| Flash Point |
413.6ºC
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| LogP |
-5.8
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| Hydrogen Bond Donor Count |
10
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| Hydrogen Bond Acceptor Count |
14
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
34
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| Complexity |
681
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| Defined Atom Stereocenter Count |
15
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| SMILES |
CN[C@@H]1[C@@H](O)[C@@H](O[C@H]2[C@H](N)C[C@H](N)[C@@H](O[C@H]3O[C@H]([C@H](O)C)[C@@H](O)[C@H](O)[C@H]3N)[C@@H]2O)OC[C@]1(C)O
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| InChi Key |
BRZYSWJRSDMWLG-DJWUNRQOSA-N
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| InChi Code |
InChI=1S/C20H40N4O10/c1-6(25)14-11(27)10(26)9(23)18(32-14)33-15-7(21)4-8(22)16(12(15)28)34-19-13(29)17(24-3)20(2,30)5-31-19/h6-19,24-30H,4-5,21-23H2,1-3H3/t6-,7+,8-,9-,10-,11+,12+,13-,14-,15-,16+,17-,18-,19-,20+/m1/s1
|
| Chemical Name |
(2R,3R,4R,5R)-2-[(1S,2S,3R,4S,6R)-4,6-diamino-3-[(2S,3R,4R,5S,6R)-3-amino-4,5-dihydroxy-6-[(1R)-1-hydroxyethyl]oxan-2-yl]oxy-2-hydroxycyclohexyl]oxy-5-methyl-4-(methylamino)oxane-3,5-diol
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| Synonyms |
geneticin; 49863-47-0; G418; antibiotic G 418; antibiotic G-418; G-418; G 418; A08F5XTI6G;
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O: ~50 mg/mL
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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