Synonym: G 418 disulfate salt solution; O-2-Amino-2,7-dideoxy-Δ-glycero-α-Δ-glucoheptopyranosyl[1->4]-O-3-deoxy-4C-methyl-3-[methyl-amino]-β-L-arabinopyranosyl-Δ-streptamine, disulfate salt; neomycin; Antibiotic G418
CAS Number: 108321-42-2
Empirical Formula (Hill Notation): C20H40N4O10 · 2H2SO4
Molecular Weight: 692.71
MDL Number: MFCD00058314
Linear Formula: C20H40N4O10 · 2H2SO4
Product Type: Chemical
Selection of transfected cells by fluorescence microscopy Fluorescence microscopy was used to determine the percentage of PC-3 cells expressing enhanced green fluorescent protein (eGFP) seven days post transfection. Increasing selection pressure of higher G-418 concentrations (above 500 μg/mL) is accompanied by an enrichment of fluorescent cells (> 80%).
Selection of transfected cells by fluorescence microscopy In the absence of selection pressure from G-418, cultures contain only 40% fluorescent cells.
Determination of working concentration (I) Concentration-response curve. Different concentrations of G-418 solution were tested on adherent PC-3 prostate cancer cells to determine ability of the antibiotic to inhibit cell growth. The inhibitory effect is seen as early as 3 days (I) and is more pronounced at 7 days (II) after exposure. Concentrations between 250 μg/mL and 500 μg/mL seem to be sufficient to completely block the growth of non-transfected PC-3 cells
Determination of working concentration (II) Inhibition of PC-3 cells after 7 days of selection. Different concentrations of G-418 were tested as described above.
Selection of transfected cells (WST-1) After the first round of selection, the proliferation rate of non-transfected cells is strongly diminished at G-418 concentrations above 100 μg/mL, as shown by the WST-1 assay .
Selection of transfected cells (WST-1) After the first round of selection, the proliferation rate of non-transfected cells is strongly diminished at G-418 concentrations above 100 μg/mL, as shown by the WST-1 assay .
Selection of transfected cells (fluorescence) Evaluation of the percentage of PC-3 cells expressing enhanced green fluorescent protein (eGFP) after two rounds of selection by G-418 (above 500 μg/mL). Only transfected cells survived, as determined with fluorescence microscopy.
Selection of transfected cells (fluorescence) Evaluation of the percentage of PC-3 cells expressing enhanced green fluorescent protein (eGFP) after two rounds of selection by G-418 (above 500 μg/mL). Only transfected cells survived, as determined with fluorescence microscopy.
| antibiotic activity spectrum |
protozoa |
| assay |
98% (TLC) |
| concentration |
50 mg/mL |
| form |
solution |
| InChI |
1S/C20H40N4O10.2H2O4S/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;2*1-5(2,3)4/h6-19,24-30H,4-5,21-23H2,1-3H3;2*(H2,1,2,3,4)/t6?,7-,8+,9+,10+,11-,12-,13-,14+,15+,16-,17-,18+,19-,20+;;/m0../s1 |
| InChI key |
UHEPSJJJMTWUCP-NKCAIAFTSA-N |
| manufacturer/tradename |
Roche |
| mode of action |
protein synthesis | inhibits |
| mol wt |
692.7 Da |
| packaging |
pkg of 100 ml (04727894001 [5 x 20 ml, equivalent to 5 g]) |
| |
pkg of 20 mL (04727878001 [equivalent to 1 g]) |
| Quality Level |
100  |
| SMILES string |
OS(O)(=O)=O.OS(O)(=O)=O.CN[C@H]1[C@H](O)[C@@H](OC[C@@]1(C)O)O[C@H]2[C@H](N)C[C@H](N)[C@@H](O[C@H]3O[C@H]([C@H](C)O)[C@@H](O)[C@H](O)[C@H]3N)[C@@H]2O |
| storage temp. |
2-8°C |
| technique(s) |
transfection: suitable |
| Application: |
G-418 Solution is used to select eukaryotic cells that are stably transfected with the neomycin resistance gene (neo), and to maintain the phenotype (neor) of resistant cells.
G-418 is also used to eliminate contaminating fibroblasts from mixed cultures. |
| Biochem/physiol Actions: |
Mode of Action: G418 blocks polypeptide synthesis and protein elongation by inhibiting synthesis at the 70S and 80S ribosomes.
Antimicrobial Spectrum: G418 selects for cells stably transfected with an iNOS promoter construct and neomycin resistance gene and shows activity against protozoa and helminthes. |
| Features and Benefits: |
• Convenient sterile-filtered solution for immediate use – saves valuable time and effort, and avoids potential errors arising from the preparation of sterile working stock solution from G-418 powder.
• Consistent quality and potency – shows no lot-to-lot variation in its ability to select resistant cells.
• Function tested in a cell culture assay - ensures effectiveness.
• Very pure, as indicated by TLC and microbiological assays – allows you to use less antibiotic while producing greater selection pressure. Lack of contaminants means minimal toxicity.
• Cost effective, since the selected colonies are stable.
Contents
Aqueous solution of G-418, 892 μg/mg, filtered through 0.2μm pore size membrane |
| General description: |
O-2-Amino-2,7-dideoxy-δ-glycero-α-δ-glucoheptopyranosyl[1->4]-O-3-deoxy-4C-methyl-3-[methyl-amino]-β-L-arabinopyranosyl-δ-streptamine, disulfate salt G-418, an aminoglycoside antibiotic, is used as a dominant selection agent in transfection experiments. The antibiotic interferes with the function of 80S ribosomes and blocks protein synthesis in eukaryotic cells. Transfection of neomycin-resistance genes into cells results in resistance to G-418 and enables the cells to grow in culture media containing G-418. |
| Other Notes: |
For life science research only. Not for use in diagnostic procedures. |
| Preparation Note: |
Working concentration: The optimal concentration of G-418 must be determined experimentally and varies with the cell type used.
Active weight: >700μg/mg
Recommended concentration in cell culture: The optimal concentration of G-418 must be determined experimentally; it varies from 100μg/ml to 1mg/ml, depending on the cell type used.
Specific rotation: 104° to 121°, in water (on dry basis, according to USP XXII). |
| Quality: |
Purity: = 98%, as shown by TLC.
Function testing: in cell culture. |
| Purity |
98% (TLC) |
| Storage Temp. |
2-8°C |
| UNSPSC |
12352200 |
