Application

Hygromycin B antibiotic was used for hygro resistance effect in the following cells: transfected 293T cells with mixture of plasmids at 160 μg/ml. MCF-7^ALOX12 cells at 150 μg/ml.MSCV Luciferase PGK-hygro plasmid.STAT3 Reporter HeLa Stable Cell Line (stably transfected with Luciferase gene under the control of STAT3 promoter) maintained in DMEM supplemented with 10% FBS.

Hygromycin B is an aminoglycoside antibiotic isolated from Streptomyces hygroscopicus. It has been used to study protein synthesis at the level of the 70S ribosome translocation and mRNA template misreading, as an antiviral agent by selectively penetrating cells rendered permeable by virus infection and inhibiting translation, and as a selection agent for hygromycin resistance gene transformed cells. It is recommended for use as a selection agent at 100-800 µg/mL, specifically at 100 µg/mL for prokaryotes, 200 µg/mL for lower eukaryotes and 150-400 µg/mL for higher eukaryotes.

Biochem/physiol Actions

Mode of Action: The product acts by inhibiting protein synthesis by inducing the misreading of the m-RNA template in the prokaryote, with the potency to inhibit translation. Antimicrobial Spectrum: Hygromycin B acts against bacteria, fungi and higher eukaryotic cells.

Caution

Hygromycin B products should be stored as supplied at 2-8°C, and the dry solid is table for at least 5 years if stored at 2-8°C. It is stable at 37°C for 30 days. (For solutions) This solution is stable as supplied for two years if stored at 2-8°C.

General description

Chemical structure: aminoglycoside

Aminoglycosides are the most widely used class of antibiotics that target ribosome. Hygromycin B is an aminocyclitol antibiotic. The organism, S. hygroscopicus contains phosphotransferase (HPH) activity, which phosphorylates hygromycin B helping in autoimmunity against the toxic effects of the drug.

Preparation Note

This product is purified by ion exchange chromatography. Hygromycin B is soluble in water at concentrations >50 mg/mL, and also soluble in methanol or ethanol. Solutions should be sterilized by filtration, not by autoclaving. Additionally, Hygromycin B solutions have been reported to lose activity on freezing, and since they are stable refrigerated, freezing should be avoided.