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潮霉素B

CAS:No.31282-04-9

英文名称:Hygromycin B

分子式:C20H37N3O13

分子量:527.52

储存条件:Powder:2-8℃,2 years;Insolvent(母液):-20℃,6 months;-80℃,1 year

纯度:≥90%

潮霉素B
满3999赠露营车
货号:
IH0160
品牌:
Solarbio
SKU 北京总部 北京海淀 武汉 广州
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使用本产品的案例(仅供参考

In Vitro

Cell (HEK293T cells; 100-400 mg/mL Hygromycin B)

In brief, the plasmid DNA was isolated from the TOP10 F' Escherichia coli containing pcDNA3.1 /Hygro (+)-ASPH using  plasmid miniprep kit according to the manufacturer's protocol.  The plasmid was then linearized using FspI digestion. HEK293T cells (2 × 105 cells/well) were seeded in 6-well cell culture plates to give optimal confluency (70%) at the time of transfection. Transfection was carried out using 4 μg of plasmid DNA and 8 μL of transfection reagent. After three days, the medium was replaced with a fresh medium containing 100 mg/mL hygromycin B (Solarbio Science & Technology, China). The plate was fed with fresh selective medium every 4 days and was screened for resistant cells remained in different parts of the plate. The detected resistant cells were monitored for 4 weeks to spot separated subclones with the ASPH expression potential.  Five selected subclones were carefully scraped off the plate using sterile scrapers and transferred to separate flasks. After 2 days, the cells were trypsinized and redistributed in the flask. The mRNA expression level of each flask was evaluated as described below (subsection qRT-PCR) to select the subclones with the highest ASPH expression. Then, the concentration of hygromycin B was gradually increased in the selected flasks to reach 400 mg/mL after 4 weeks of culture.

来源文献:Bakhtiari H, Palizban AA, Khanahmad H, Mofid MR. An innovative cell selection approach in developing human cells overexpressing aspartyl/asparaginyl β-hydroxylase. Res Pharm Sci. 2020 Jul 3;15(3):291-299.doi: 10.4103/1735-5362.288436.


备注:
以上数据均来自公开文献, Solarbio暂未进行独立验证, 仅供参考。
These protocols are for reference only. Solarbio does not independently validate these methods.

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