Puromycin

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

Puromycin is a selection antibiotic for mammalian, insect, and bacterial cells, available as a sterile, cell culture-tested solution. Puromycin dihydrochloride is an aminonucleoside antibiotic naturally produced by Streptomyces alboniger as a tyrosyl tRNA analog.

Puromycin specifically inhibits peptidyl transfer on both prokaryotic and eukaryotic ribosomes. This antibiotic is highly active on Gram-positive bacteria and mammalian and insect cells but is weakly active on Gram-negative bacteria.

Resistance to puromycin is conferred by the Pac gene encoding a puromycin N-acetyl-transferase isolated from a Streptomyces strain [1]. Vectors carrying only one drug resistance marker for selection can be designed with the Pac gene.

The recommended working concentrations are 0.5 - 10 μg/ml for selection in mammalian cells and 100 - 125 μg/ml for selection in E. coli.

Key features

• Produced in our own facilities
• Each lot thoroughly tested to ensure lot-to-lot reproducibility
• Potency validated in blasticidin-sensitive and blasticidin-resistant mammalian cell lines
• Absence of long-term effects confirmed in blasticidin-resistant cells

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

Reference:

1. Lacalle R. et al., 1989. Molecular analysis of the pac gene encoding a puromycin N-acetyl transferase from Streptomyces alboniger. Gene. 79:375-80. 

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Specifications

CAS number: 58-58-2

Formula: C₂₂H₂₉N₇O₅ • 2HCl

Molecular weight: 544.3 g/mol

Appearance (form): Liquid

Appearance (color): Colorless

Product concentration: 10 mg/ml 

Buffer: HEPES

pH: 6.0 - 7.0

Purity: ≥ 98% (HPLC)

Sterility: 0.2 um filter sterilized

Endotoxin level: < 5 EU/mg

Working concentration: 0.5 - 10 μg/ml (mammalian cells), 100 - 125 μg/ml (E. coli)

Quality control: Cell-culture tested

Structure :

Contents

Puromycin dihydrochloride is supplied as a sterile filtered solution at 10 mg/ml in HEPES buffer.

This product is available in three pack sizes:

• ant-pr-1: 10 x 1 ml (100 mg)
• ant-pr-5: 50 x 1 ml (500 mg)
• ant-pr-5b: 1 x 50 ml (500 mg)

Puromycin is shipped at room temperature.

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

Puromycin is a harmful compound. Refer to safety data sheet for handling instructions.

FAQs

What is the best way to resuspend Puromycin?
We recommend to resuspend Puromycin in HEPES buffer (5 mM, pH 7.3)

We have noticed a crystalline precipitate of Puromycin when thawing a vial left at -20°C, is this normal?
Yes, during storage a crystalline precipitate of Puromycin may form. If this occurs, heat the product at 37°C for 30 minutes until the crystalline precipitate disappears. The formation of a crystalline precipitate does not affect the activity of the product.

What is the preferred storage method for Puromycin?
Puromycin 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. 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 Puromycin in mammalian cells?
The working concentration of Puromycin in mammalian cell lines varies between 1-10 μg/ml. In an initial experiment we recommend to determine the optimal concentration of puromycin required to kill your host cell line. Please note that Puromycin quickly kills eukaryotic cells that do not contain the pac gene.
For more detailed information, please refer to the technical data sheet for this product.

What is the selection concentration of Puromycin in E.coli?
Puromycin has low activity in E. coli. However, puromycin-resistant transformants can be selected in LB agar medium, with the pH adjusted to 7.5-8, supplemented with 100-125 μg/ml of Puromycin.

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