Phleomycin

Selection antibiotic for bacteria and mammalian cells – Endotoxin-tested

Phleomycin, CAS 11006-33-0, is a selection antibiotic for mammalian and bacterial cells. It is sterile, cell culture tested, and available as a solution. This member of the bleomycin family is a glycopeptide antibiotic isolated from a mutant strain of Streptomyces verticillus.

Phleomycin binds and intercalates DNA thus destroying the integrity of the double helix [1]. It is active against most bacteria, filamentous fungi, yeast, plant and animal cells.

Use of phleomycin is recommended for cells poorly sensitive to Zeocin®, i.e. filamentous fungi and some yeasts. Phleomycin resistance is conferred by the Sh ble gene from Streptoalloteichus hindustanus, which encodes a protein that binds to phleomycin, inhibiting its DNA cleavage activity.

Typically, phleomycin is used at a concentration of 10 µg/ml for yeasts and 25 - 150 µg/ml for filamentous fungi.

Key features

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

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

Reference:

1. Wilson JB, et al., 2010. Several tetratricopeptide repeat (TPR) motifs of FANCG are required for assembly of the BRCA2/D1-D2-G-X3 complex, FANCD2 monoubiquitylation and phleomycin resistance. Mutat Res. 689(1-2):12-20.

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Specifications

CAS number: 11006-33-0

Appearance (form): Liquid or powder

Appearance (color): Blue

Product concentration: 20 mg/ml (solution)

Buffer: HEPES

pH: 7.1-7.7 (solution)

Sterility: 0.2 um filter sterilized

Endotoxin level: < 1 EU/mg

Working concentration: 10 μg/ml (yeasts), 25 - 150 μg/ml (filamentous fungi)

Quality control: Cell-culture tested

Contents

Phleomycin is supplied as a sterile filtered blue solution at 20 mg/ml in HEPES buffer.

This product is available in three pack sizes:

• ant-ph-1: 5 x 1 ml (100 mg)
• ant-ph-5: 25 x 1 ml (500 mg)

Phleomycin is also supplied as a blue powder:

• ant-ph-2p: 250 mg
• ant-ph-5p: 2 x 250 mg (500 mg)

Phleomycin is shipped at room temperature.

Upon receipt it should be stored at 4°C or -20°C.

FAQs

What is the preferred storage method for Phleomycin?
Phleomycin is shipped at room temperature. Upon receipt, it should be stored at 4°C for short term storage or -20°C for long term storage. Avoid repeated freeze-thaw cycles. The expiry date is specified on the product label.
For Phleomycin powder, once resuspended at 20 mg/ml, it can be stored at 4°C for 12 months or -20 °C for 18 months.
For Phleomycin solution, 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.

What is the selection concentration of Phleomycin in mammalian cells?
The working concentration of Phleomycin for mammalian cell lines varies between 5 -50 μg/ml. In an initial experiment we recommend to determine the optimal concentration of Phleomycin required to kill your host cell line.
For more detailed information, please refer to the technical data sheet for this product.

What is the selection concentration of phleomycin in E.coli?
Phleomycin-resistant transformants are selected in low salt LB agar medium supplemented with 5 μg/ml of Phleomycin. Notably, common E. coli recipient strains (i.e. HB101, DH5α, MC1061) transformed with vectors carrying bleomycin resistant genes, such as Sh ble and Tn5, are resistant to Phleomycin.

Any questions about our Selection antibiotics ? Visit our frequently asked questions page