Jump to navigation. G also known as G, G sulfate or Geneticin is an aminoglycoside antibiotic produced by Micromonospora rhodorangea. G blocks polypeptide synthesis by inhibiting the elongation step in both prokaryotic and eukaryotic cells. Quality Control: Each lot is thoroughly tested to ensure the absence of lot-to-lot variation.
Cell-culture tested: potency validated in Gsensitive and Gresistant mammalian cell lines. Non-cytotoxicity of trace contaminants: absence of long-term effects confirmed in Gresistant cells. Formula: C 20 H 40 N 4 O G is shipped at room temperature. However, the optimal concentration needs to be determined for your cells. Suggested concentrations of G for selection in some examples of mammalian cells are listed below:. Arikawa K. Ligand-dependent inhibition of B16 melanoma cell migration and invasion via endogenous S1P2 G protein-coupled receptor.
Requirement of inhibition of cellular RAC activity. J Biol Chem. DiLillo DJ. J Immunol. Figueroa K. Selectivity of agonists for the active state of M1 to M4 muscarinic receptor subtypes. J Pharmacol Exp Ther. Kwon JA. Hermoso M. Cell volume regulation in response to hypotonicity is impaired in HeLa cells expressing a protein kinase Calpha mutant lacking kinase activity. Hennen S. Decoding signaling and function of the orphan G protein-coupled receptor GPR17 with a small-molecule agonist.
Chen Q. Expression of Drosophila trehalose-phosphate synthase in HEK cells increases hypoxia tolerance. Tukhvatulin A. Due to translation G inhibited polypeptide synthesis and protein elongation. G binds to the 80S subunit of the ribosome by blocking the elongation phase of protein synthesis, both in prokaryotic and eukaryotic cells. It also shows activity against protozoa, cecal amoebiasis, and helminths. G inhibits protein synthesis in prokaryotic and eukaryotic cells.
It is toxic to bacteria, yeast, protozoans, higher plant, and mammalian cells. The working concentration for the purpose of selection varies with cell type, media, growth conditions and cell metabolic rate. Below are recommended G concentrations based on cell type. G and Geneticin have the same chemical structure. G is the generic chemical while Geneticin is a registered trademark brand of LifeTech Corporation.
Small lot-to-lot variances do occur. This gene allows production of aminoglycoside 3-phosphotransferase, which inactivates G, neomycin , and kanamycin via phosphorylation.
Cells can escape selection if the antibiotic is used at a very low concentration or if the cell density on the plate is too high.
Additionally, cells rapidly proliferating are killed faster than those only slowly proliferating. Control cells should die within days after addition of the antibiotic allowing colonies of resistant cells to form by days.
Yes, cells can continue to divide up to two times in the presence of lethal doses of the antibiotic. The effect of the product usually becomes apparent within two days. To convert the potency reported on the powder to liquid potency, divide the potency by and multiply this by the desired potency. We recommend setting a dose response curve for both sensitive and resistant mammalian cells.
These concentrations are reported as dry weight. G is most effective against dividing cells. By permitting cell growth prior to the addition of G, 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.
Test a range of concentrations at least 6 to ensure that you determine the minimum concentration necessary for your cell line. Using an analytical balance, weigh an appropriate amount of G powder use formula below. Reconstitute aseptically by adding the powder to distilled water pH 5. Potency is always measured against a reference by the inhibition ring method.
The inhibitory effect was measured on B. JavaScript seems to be disabled in your browser. For the best experience on our site, be sure to turn on Javascript in your browser. About G Sulfate G Sulfate also known commercially as Geneticin is an aminoglycoside antibiotic that is synthesized from Micromonospora rhodorangea and functions by causing a chain of nonsense mutations during translation.
What is the working concentration for selection? Show 5 10 20 50 Add to Cart. Posted By: AG Blogger.
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