Hygromycin B Gold

Selective antibiotic for the hph gene

ABOUT

Selection antibiotic for bacteria and mammalian cells – Sterile, endotoxin-tested

Hygromycin B Gold is an ultrapure Hygromycin B used for the selection of bacteria and eukaryotic cells, provided as a sterile, cell culture-tested solution. Hygromycin B is an aminoglycoside antibiotic produced by Streptomyces hygroscopicus, active against bacteria, fungi, and higher eukaryotic cells. It interferes with translocation [1] causing mistranslation of the 70s ribosomal subunit [2], thus inhibiting protein synthesis.

Hygromycin B is used as a selective agent on a wide variety of eukaryotic and prokaryotic cells expressing the hph resistance gene [3-5]. Due to its unusual mechanism of action, Hygromycin B can be used for dual-selection experiments. Since Hygromycin B is active in both bacteria and mammalian cell lines, vectors can be designed that carry only one drug resistance marker for selection. Selection with Hygromycin B Gold is typically achieved using concentrations ranging from 50 to 200 µg/ml in mammalian cells and 50 to 100 µg/ml in bacteria.

 

Key features 

  • Each lot is thoroughly tested to ensure lot-to-lot reproducibility.
  • Potency validated in hygromycin-sensitive and hygromycin-resistant mammalian cell lines
  • Absence of long-term effects confirmed in hygromycin-resistant cells

 

References:

1. Cabanas M. et al., 1978. Dual interference of Hygromycin B with ribosomal translocation and with Aminoacyl-tRNA recognition. Eur. J. Biochem. 87:21‑7.
2. Gonzales, A. et al., 1978. Studies on the mode of action of hygromycin B, an inhibitor of translocation in eukaryotes. Biochem Biophys Acta 521:459‑69.
3. Gritz L. & Davies J., 1983. Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae. Gene 25:179-88.
4. Cullen D. et al., 1987. Transformation of Aspergillus nidulans with the hygromycin-resistance gene, hph. Gene 57:21-6.
5. Santerre R. et al., 1984. Expression of prokaryotic genes for hygromycin B and G418 resistance as dominant-selection markers in mouse L cells. Gene 30:147-56.

All InvivoGen products are for internal research use only, and not for human or veterinary use.

SPECIFICATIONS

Specifications

Product concentration

100 mg/ml

Applications

Eukaryotic selection/stable cell line generation (tested), bacterial selection (tested)

CAS number
31282-04-9
Chemical formula

C20H37N3O13, HCl

Molecular weight
563.5 g/mol
Purity
≥ 90% (HPLC)
Buffer

HEPES

pH

7.5 - 8.1

Appearance (form)
Liquid
Appearance (color)
Colorless to light brown
Working concentration

50 - 200 µg/ml (mammalian cells), 50 - 100 µg/ml (E.coli)

Sterility

0.2 µm filtration

Endotoxin

< 0.5 EU/mg

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    Hygromycin B Gold (100 mg/ml solution)
  • Cat code: 
    ant-hg-1
  • Quantity: 
    1 g (10 x 1 ml)

Shipping & Storage

  • Shipping method:  Room temperature

    • Storage:

    • 4°C or -20°C
    Stability: The product is stable for 3 months at room temperature.

    Caution:

    • Avoid repeated freeze-thaw cycles
    • This product is a harmful compound. Refer to SDS for handling instructions

DOCUMENTS

Documents

Hygromycin B Gold (100 mg/ml solution)

Technical Data Sheet

Safety Data Sheet

Certificate of analysis

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FAQ

Hygromycin B Gold

Hygromycin B Gold is shipped at room temperature.
Upon receipt, it should be stored at 4°C or -20°C.
Avoid repeated freeze-thaw cycles.
The expiry date is specified on the product label. Once opened it remains stable for 1 year at -20°C providing you avoid repeated freeze-thaw cycles and it can be kept at 4°C for 3 months provided it is kept in sterile conditions.

The working concentrations of Hygromycin B Gold for mammalian cell lines vary from 50 to 200 μg/ml; in a few cases, up to 500 μg/ml. In a starting experiment we recommend to determine the optimal concentration of Hygromycin B Gold required to kill your host cell line. In general, Hygromycin B Gold is used at a concentration of 200 μg/ml, a 500-fold dilution from the stock solution. 
For more detailed information, please refer to the technical data sheet for this product.

Hygromycin-resistant transformants are selected in low-salt LB agar medium (yeast extract 5g/l, tryptone 10 g/l, NaCl 5 g/l, agar 15 g/l, pH 8) supplemented with 50 to 100 μg/ml of Hygromycin B Gold.
Plates containing Hygromycin B Gold are stable for 1 month when stored at 4 °C.

We recommend to use Hygromycin B Gold at a concentration of 100 – 150 µg/ml for high salt LB conditions

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