What is the concentration of G418 for selection?
What is the concentration of G418 for selection?
400 to 1000 µg/mL
The concentration range of G418 that is commonly used ranges from 400 to 1000 µg/mL for selection of mammalian cells and 200 to 300 µg/mL for maintenance.
How do you dilute G418?
200 mg/ml G-418 Stock Solution:
- Weight 2gr of G418 (CAS: 108321-42-2 mw=692.71).
- Add 10ml of sterile H2O.
- Prewet a 0.22um syringe filter by drawing through 5-10ml pf sterile H2O and discard water.
- Sterilize G418 Stock through through the prepared 0.22um syringe filter.
- Stock may be kept at -20C for 1 year.
How does geneticin inhibit cell growth?
G-418, otherwise known as Geneticin®, is produced by Micromonospora rhodorangea through fermentation processes. And it works by blocking polypeptide synthesis by preventing the elongation step within eukaryotic cells. plant cells and the NEO gene in mammalian cells, which encodes aminoglycoside 3′-phosphotransferase.
How long is G418 selection?
3 to 7 days
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.
How do I reconstitute G418?
Stock Solution Guideline: Reconstitute aseptically by adding the powder to distilled water (pH 5.6 to 7.0) to the desired concentration and sterile filter. NOTE: For G418, units/mg is exchangeable with µg/mg.
Can I use G418 instead of neomycin?
Bottom line: Use G418 for single-selection experiments on eukaryotic cells. Use neomycin or kanamycin for single-selection experiments on prokaryotic cells where you are selecting for neomycin resistance genes.
Is geneticin same as gentamicin?
Geneticin is a 2-deoxystreptamine antibiotic that is structurally related to gentamicin. Although not used clinically, it is employed as an antibiotic in molecular genetics experiments. Geneticin is toxic to bacteria, yeast, higher plant and mammalian cells, protozoans, and helminths.
Can you freeze G418?
G418 is shipped at room temperature. Upon receipt, it should be stored at 4°C for short term storage or -20°C for long term storage. Avoid repeated freeze-thaw cycles.
Is gentamicin the same as G418?
Commonly know as G418 or G-418, Gibco Geneticin reagent is an aminoglycoside related to gentamicin, and is commonly used as a selective agent for eukaryotic cells. As an analog of neomycin sulfate, it interferes with the function of 80S ribosomes and protein synthesis in eukaryotic cells.
What is the best concentration of G418 to use?
Concentration of G418: you should use the LOWEST concentration as possible, i.e. the concentration which basically will kill all control cells within a week or two (not faster). You do not want to have all your cells dead after 48h, it is not a good point.
Should G418 be added before or after cell growth?
By permitting cell growth prior to the addition of G418, one will maximize the selection characteristics of the antibiotic for both sensitive and resistant cells. To generate a stable cell line expressing your protein of interest, you need to determine the minimum concentration of antibiotic required to kill your untransfected host cell line.
How do you use seselection G418 sulfate?
Selection procedure for mammalian cells G418 sulfate is normally used at a concentration of 400 μg/ml. After transfection with a plasmid containing the neo gene, cells are incubated in their regular growth medium containing G418 to select for stable transfectants.
What is the recommended concentration of G418 for a kill curve assay?
It is recommended that a kill curve assay with various concentrations of G418 be performed. The concentration range of G418 that is commonly used ranges from 400 to 1000 µg/mL for selection of mammalian cells and 200 to 300 µg/mL for maintenance.
