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- 保质期：
1 g (10 x 1ml)
- CAS号：
ANT-GN-1
- 英文名：
G418 (Geneticin)
- 保存条件：
低温
- 供应商：
G418 (solution)
- 库存：
G418 (solution)
- 规格：
1 g (10 x 1ml)
G418 (Geneticin)
去除内毒素的基因筛选抗生素
Selection antibiotic: cell culture tested, sterile reagent
G418 (Geneticin)
Selection antibiotic: cell culture tested, sterile reagent
G418, also known as G418 sulfate and Geneticin, is an aminoglycoside antibiotic similar in structure to gentamicin B1, produced by Micromonospora rhodorangea. G418 blocks polypeptide synthesis by inhibiting the elongation step in both prokaryotic and eukaryotic cells. 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 three to seven days with concentrations ranging from 400 to 1000 µg/ml. Cells that are dividing are affected sooner than those that are not.\
Product concentration: 100 mg/ml
CAS number: 108321-42-2
Quality Control: Each lot is thoroughly tested to ensure the absence of lot-to-lot variation.
Purity: ≥ 90% (HPLC)
Endotoxin level: < 0.5 EU/mg
Physicochemical characterization: pH, appearance
Cell-culture tested: potency validated in G418-sensitive and G418-resistant mammalian cell lines
Non-cytotoxicity of trace contaminants: absence of long-term effects confirmed in G418-resistant cells
G418 is supplied as a sterile filtered solution at 100 mg/ml in HEPES buffer. This product is available in two pack sizes:
G418规格- ant-gn-1: 10 x 1 ml (1 g)
- ant-gn-5: 1 x 50 ml (5 g)
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文献和实验Citations
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（1）DMSO广泛应用在人、动物细胞株及细菌噬菌体λ的低温防护中，制备DMSO溶液用来冷冻细胞的步骤如下： 1)配制冷冻培养基：含有细胞培养用培养基、10-20% 血清和5-10% DMSO； 2)用胰酶或其他合适的方法消化贴壁细胞；（细胞最好处于对数增长期） 3)低速离心（4 °C，250 × g, 10min）收集细胞，吸去培养基； 4)用冷冻培养基悬浮细胞，使细胞密度为106 -107 cells/ml；
培养基)，每个浓度4个重复，则每次换液需各个浓度培养基4ml，现用现配。 3.细胞培养： 将冻存的细胞复苏培养，，传代3至4次使细胞达到良好的生长状态,铺24孔板(20000/孔)，12h换液，加筛选培养基培养。 4.确定G418最佳筛选浓度： 将培养孔中培养基吸除，PBS洗涤一次，每孔加入不同浓度筛选培养基，隔天更换一次筛选培养基，培养10-14天，以最低细胞全部死亡浓度为基准。 注：实验中所确定的最佳筛选浓度为1200ug/ml。 5.铺6孔
不得的，还是要踏踏实实的做，摸索方法还是很简单的：将细胞稀释至1000cell/ml，每孔100μl加入有培养基的24孔板，将每孔中的G418浓度稀释至0,，100，200，300，400，500，600，700，800，900，1000，1100μg/ml等12个级别，培养10-14天，以最低细胞全部死亡浓度为基准，一般400-800左右，筛选时比该浓度再高一个级别，维持使用筛选浓度的一半。不同的质粒转染同一种细胞，筛选的G418浓度是一样的，但是否都能筛选得到稳定株就很难说了，祝你好运！
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