High quality Hygromycin B with best price CAS NO.31282-04-9 CAS NO.31282-04-9

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1 piece=100mg

Hygromycin B
CAS:31282-04-9
Molecular formula: C20H37N3O13
Molecular weight: 527.52
Purity: >90%
Chemical properties: White to white powder, soluble in water, methanol and ethanol.
Uses: Biochemical research
Mechanism of action: Hygromycin B is an aminoglycoside antibiotic metabolized by Streptomyces hygroscopicus, which kills bacteria, fungi, and higher eukaryotic cells by inhibiting protein synthesis. Hydomycin B inhibits protein synthesis by interfering with 70S ribosome translocation and inducing misreading of the mRNA template. Currently, it is commonly used to screen and maintain prokaryotic or eukaryotic cells containing hygromycin resistance genes.
Resistance genes:
Resistance to hygromycin B is derived from the gene encoding hygromycin B phosphotransferase (Hph). Two sources have been found so far:
? Hph resistance gene produced by Streptomyces hygroscopicus;
? Hph resistance gene carried by Escherichia coli, Klebsiella pneumoniae (commonly used);
Biological Applications:
1) Screening and maintenance of cultured prokaryotic or eukaryotic cells (plant cells and mammalian cells) stably transfected with Hph+ vectors;
2) Due to differences in mode of action, it is often used in combination with G418 (GeneticinTM), Zeocin™ and blasticidin S to screen cell lines stably transfected with two different vectors;
3) Antiviral agents, because hygromycin B selectively penetrates into cells that enhance permeability due to viral infection, and has the effect of inhibiting translation;
4) Add to animal feed as dewormer;
Application concentration:
The working concentration of hygromycin B for screening stable strains needs to vary according to cell type, medium, growth conditions, and cell metabolic rate. The recommended concentration is 50-1000 μg/mL, and the optimal concentration needs to be determined by the kill curve.
Mammalian cells ·200-500 μg/mL; Bacterial/plant cells ·100-300 μg/mL; Fungi ·300-1000 μg/mL;
Product usage:
Use one: Kill curve to determine the best kill concentration (for reference only, varies by individual test system)
(1) Untransfected cells were added into 96-well plates at tissue culture level, and the cell density was about 50-200 cells/well; After the cells were attached to the wall, 200μL fresh medium containing different concentrations (e.g. 50-1000μg/mL, at least 5 concentrations) of hygromycin B was added to each well.
(2) Cells were incubated at 37℃ in 5% CO2 incubator for 10-14 days;
(3) After 5-7 days of culture, replace the new culture medium with the corresponding concentration of hygromycin B;
(4) Cell viability was assessed by cell proliferation methods such as MTT and CCK-8 after 10-14 days; Toxic effects can also be determined by examining the number of cell clones or the percentage confluence rate. Generally, the minimum concentration that can kill all cells in 10-14 days is selected as the optimal screening concentration.
Use two: Screen stable transfected cells
(1) For adherent cells, the transfected cell medium in a 60mm petri dish was sucked directly, and then 5-6ml fresh medium containing hygromycin B was added; For suspended cells, aseptic conditions were 250x g, centrifugation was performed for 10min to remove the old transfected cell medium, and then suspended in about 5ml of fresh medium containing hygromycin B;
(2) After 5-7 days, replace the fresh medium containing hygromycin B by the above method;
(3) Incubate the cells again for 5-7 days;
(4) After 10-14 days of incubation, the culture system contained only living cells capable of expressing hygromycin B resistant phenotype. Therefore, after screening, as in step 1, replace the fresh medium without hygromycin B.

Nanjing Duly Biotech Co.,Ltd 

has been established for 11 years. It  is mainly engaded in biological and chemical reagents,including enzyme reagents, proteins, nucleic acids, pigments, amino acids, vitamins,carbohydrates, buffers, surfactants,culture media and Separation of materials and kits.There are more than 10,000 products in total.

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