Description
G418 Sulfate from GoldBio is a high-purity aminoglycoside antibiotic widely used for the selection of eukaryotic and yeast cells that express the neomycin resistance gene (neoR).
With ≥98% purity and exceptional lot-to-lot consistency, this product is ideal for generating stable transfectants in mammalian cell lines, yeast systems, and embryonic stem cells for gene editing applications.
Its robust cytotoxic activity enables precise screening of transgenic constructs, shRNA vectors, and CRISPR integrations. Whether developing stable cell lines for protein production or executing high-throughput compound screening, GoldBio’s G418 ensures reliable performance and reproducible results.
G-418 functions by causing a chain of nonsense mutations during translation, ultimately resulting in inhibition of polypeptide synthesis and protein elongation. Resistance to G-418 is conferred through the nptll gene in plant cells and the neo gene in mammalian cells, which encode aminoglycoside 3'-phosphotransferase. For gene selection applications, a kill curve can be made to determine the minimum effective G418 concentration to kill non-resistant cells. G418 disulfate is freely soluble in water.
This product has been tested against both sensitive and resistant cells at Gold Biotechnology Labs.
Common Research Applications
(Click each for more information)
Selection of Stably Transfected Eukaryotic Cells
-
Purpose: To select mammalian and other eukaryotic cells stably transfected with plasmids containing a neomycin resistance gene (neoR).
-
How It Works: G418 inhibits protein synthesis by binding to the 80S ribosome, leading to cell death. Cells expressing aminoglycoside 3'-phosphotransferase (neoR) detoxify the antibiotic and survive.
-
Applications: Stable transgene expression, biopharmaceutical production cell line generation, and cell engineering workflows.
Southern, P. J., & Berg, P. (1982). Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. Journal of Molecular and Applied Genetics, 1(4), 327–341.
Yeast Genetic Manipulation and Selection
-
Purpose: To select genetically modified yeast strains carrying resistance cassettes such as kanMX.
-
How It Works: The kanMX gene encodes aminoglycoside phosphotransferase, which inactivates G418 and allows transformed cells to survive.
-
Applications: Gene deletion libraries, overexpression screens, and CRISPR-based genome editing in Saccharomyces cerevisiae and related fungi.
Wach, A., et al. (1994). New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae. Yeast, 10(13), 1793–1808.
Stable Expression of shRNA or CRISPR Constructs
-
Purpose: To enable long-term expression of gene-silencing or gene-editing tools such as shRNA and CRISPR systems.
-
How It Works: Vectors encoding Cas9, shRNA, or CRISPR components are co-expressed with neoR, allowing G418 to select for cells with stable genomic integration.
-
Applications: Functional genomics, pooled CRISPR screens, and mechanistic pathway studies.
Stewart, S. A., et al. (2003). Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA, 9(4), 493–501.
High-Throughput Screening Using Stable Cell Lines
-
Purpose: To establish stable reporter cell lines for high-throughput screening (HTS) applications.
-
How It Works: Reporter constructs (e.g., luciferase, GFP) integrated with neoR allow clonal selection using G418, ensuring consistent assay performance.
-
Applications: Drug discovery, target validation, pathway profiling, and phenotypic screening.
Inglese, J., et al. (2007). High-throughput screening assays for the identification of chemical probes. Nature Chemical Biology, 3(8), 466–479.
Generation of Transgenic Animal Models via ES Cell Selection
-
Purpose: To select embryonic stem (ES) cells that have undergone homologous recombination with neoR-containing constructs.
-
How It Works: Targeted ES cells survive G418 selection due to integration of resistance cassettes, enabling isolation of correctly modified clones.
-
Applications: Gene knockout/knock-in model development and complex transgenic animal generation.
Thomas, K. R., & Capecchi, M. R. (1987). Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell, 51(3), 503–512.
Key Benefits
- Efficient Cell Line Selection: Enables rapid and reliable isolation of stably transfected cells, accelerating downstream experimental timelines.
- Broad Host Compatibility: Active in both mammalian and yeast cells, allowing cross-system workflows with a single reagent.
- High Potency and Consistency: GoldBio’s formulation offers robust and reproducible cytotoxicity, ensuring tight selection.
- Reduces Clonal Variability: Promotes homogenous populations when used in stable expression systems.
- Supports CRISPR and Genomic Engineering: Compatible with modern gene editing vectors that use the neoR selection cassette.
Storage/Handling:
Store desiccated at -20°C.
Product Specifications:
Molecular Formula: C20H40N4O10 · 2H2SO4
Molecular Weight: 692.71 g/mol
PubChem Chemical ID: 16760463
Powered by Bioz
