G-418 Solution
ROCHE/G418-RO - solution, =98% (TLC), suitable for transfection
Synonym: G 418 disulfate salt solution; O-
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- |
| InChI key | UHEPSJJJMTWUCP-NKCAIAFTSA |
| 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.C |
| 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-g |
| 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. |
| Symbol |  GHS08 |
| Signal word | Danger |
| Hazard statements | H317 - H334 |
| Precautionary statements | P261 - P280 - P284 - P304 + P340 - P333 + P313 - P342 + P311 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | No data available |
| Flash Point(C) | No data available |
| Purity | 98% (TLC) |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352200 |