Application

Puromycin dihydrochloride from Streptomyces alboniger has been used:in the preparation of puromycin stock solution for puromycin sensitivity assay.to select lentivirus-infected cells containing puromycin resistant selection marker.

Used as a selective agent for cells that contain the resistance gene puromycin N-acetyl-transferase (PAC). Recommended for use at a range of 1-10 µg/ml. Sterilize stock solution by filtration using 0.22 µm filter then store in aliquots at −20 °C.

Puromycin dihydrochloride is a part of the amino-nucleoside family of antibiotics and is derived from Streptomyces alboniger. It is a broad spectrum antibiotic with antitumor activity, as an inhibitor of protein synthesis and has been used to study transcription regulatory mechanisms that control the sequential and coordinate expression of genes during cell differentiation. The recommended working concentration in eukaryotic cell culture is 1-10 µg/mL.

Biochem/physiol Actions

Mode of Action: Puromycin inhibits protein synthesis by causing premature chain termination acting as an analog of the 3′-terminal end of the aminoacyl-tRNA.Mode of Resistance: Puromycin acetyltransferase is an effective resistance gene. Antimicrobial Spectrum: This product is active against gram-positive microorganisms, less active against acid-fast bacilli and more weakly active against gram-negative microorganisms. Puromycin can prevent growth of bacteria, protozoa, algae and mammalian cells and acts quickly, killing 99% of cells within 2 days.

Puromycin associates in a non-specific manner with growing polypeptide chains, and thus, leads to premature termination of translation. It inhibits protein synthesis in two ways. It associates with the donor substrate, peptidyl-tRNA, in the P site and thus, functions as an acceptor substrate. Second, it competes with aminoacyl tRNA to bind with the A′ site within the peptidyl transferase center.

Caution

As supplied, this product remains active for four years when stored at -20°C.

General description

Chemical structure: peptidyl nucleoside

Puromycin is a broad spectrum antibiotic produced by Streptomyces alboniger. It is an analog of the 3′ end of aminoacyl-tRNA. It is an inhibitor of peptidyl transferase and is active in both prokaryotes and eukaryotes.

Other Notes

Keep container tightly closed in a dry and well-ventilated place.Keep in a dry place

Packaging

10, 25, 100 mg in glass bottle

Preparation Note

This product is soluble in water at 50 mg/mL. A stock solution should be prepared by filtration using a 0.22 µm filter and then stored in aliquots at -20°C. This product is also soluble in methanol at 10 mg/mL.