G-418 disulfate (Geneticin sulfate), is an aminoglycoside antibiotic, inhibits protein synthesis in eukaryotes and prokaryotes. G-418 disulfate is commonly used as a selective agent for eukaryotic cells[1].
体外研究 (In Vitro)
G418 sulfate, an aminoglycoside neomycin analogue, interferes with protein synthesis and has been used extensively for selection of mammalian cell lines that possess neomycin resistance (NR). The neomycin resistance (neo) gene is frequently used in eukaryotic vectors as a dominant selectable gene. G418 can be covalently bound to asialoorosomucoid (AsOR) to form a conjugate for hepatocyte-specific targeting and toxicity[2]. The human GD3 synthase cDNA was transfected into MDA-MB231 cells, and G-418 bulk selection was used to select cells stably overexpressing the GD3 synthase[2]. An aminoglycoside antibiotic, G418, has been shown to be an inhibitor of many pro- and eukaryotes at concentrations from 1-300 microgram/ml[4].
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量
692.71
Formula
C20H44N4O18S2
CAS 号
108321-42-2
中文名称
遗传霉素硫酸盐
运输条件
Room temperature in continental US; may vary elsewhere.
储存方式
4deg;C, stored under nitrogen, away from moisture
*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (stored under nitrogen, away from moisture)
溶解性数据
In Vitro:;
H2O : 125 mg/mL (180.45 mM; Need ultrasonic)
配制储备液
浓度溶剂体积质量
1 mg
5 mg
10 mg
1 mM
1.4436 mL
7.2180 mL
14.4361 mL
5 mM
0.2887 mL
1.4436 mL
2.8872 mL
10 mM
0.1444 mL
0.7218 mL
1.4436 mL
*
请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效。 储备液的保存方式和期限:-80°C, 6 months; -20°C, 1 month (stored under nitrogen, away from moisture)。-80°C 储存时,请在 6 个月内使用,-20°C 储存时,请在 1 个月内使用。
In Vivo:
请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液,再依次添加助溶剂:
Solubility: 50 mg/mL (72.18 mM); Clear solution; Need ultrasonic
*以上所有助溶剂都可在 MCE 网站选购。
参考文献
[1]. Giordano-Santini R, et al. An antibiotic selection marker for nematode transgenesis. Nat Methods. 2010;7(9):721-723.
[2]. Volarevic M, et al. A novel G418 conjugate results in targeted selection of genetically protected hepatocytes without bystander toxicity. Bioconjug Chem. 2007;18(6):1965-1971.
[3]. Kwon KM, et al. Disialyl GD2 ganglioside suppresses ICAM-1-mediated invasiveness in human breast cancer MDA-MB231 cells. Int J Biol Sci. 2017;13(3):265-275. Published 2017 Feb 12.
[4]. Davies J, et al. A new selective agent for eukaryotic cloning vectors. Am J Trop Med Hyg. 1980;29(5 Suppl):1089-1092.
Animal Administration [3]
To characterize the sensitivity of the trypanosome populations to G418 in vivo, bloodstream forms of T. brucei brucei GUTat 3.1 and T. brucei brucei GUTat 3.1/BBR3 are expanded separately in sublethally irradiated mice. Prior to the first peak of parasitemia, trypanosomes are collected, and aliquots containing 106 trypanosomes are inoculated intraperitoneally into mice. Twenty-four hours following infection, the mice are divided into groups and treated with G418 at a dose of 10, 20, 30, 40, 50, or 80 mg/kg of body weight (bw) by inoculating intraperitoneally 0.2 mL of the drug in sterile water. At 24 and 48 h following the first treatment, G418 is administered to animals in each group at the same dose as before, resulting in three treatments per mouse. Repeated drug treatments are necessary to ensure complete elimination of nontransfected GUTat 3.1 parasites from the mice. Mice are then monitored daily, for 33 days, for the presence of parasites by microscopic examination of wet-blood films. Animals found to be parasitemic are recorded and then removed from the experiment.
MCE has not independently confirmed the accuracy of these methods. They are for reference only.
参考文献
[1]. Giordano-Santini R, et al. An antibiotic selection marker for nematode transgenesis. Nat Methods. 2010;7(9):721-723.
[2]. Volarevic M, et al. A novel G418 conjugate results in targeted selection of genetically protected hepatocytes without bystander toxicity. Bioconjug Chem. 2007;18(6):1965-1971.
[3]. Kwon KM, et al. Disialyl GD2 ganglioside suppresses ICAM-1-mediated invasiveness in human breast cancer MDA-MB231 cells. Int J Biol Sci. 2017;13(3):265-275. Published 2017 Feb 12.
[4]. Davies J, et al. A new selective agent for eukaryotic cloning vectors. Am J Trop Med Hyg. 1980;29(5 Suppl):1089-1092.