Puromycin

SKU: orb1672963

Description

Extracted from Streptomyces alboniger bacteria, Puromycin is an aminonucleoside antibiotic that inhibits protein synthesis by interfering with the translation process. It specifically inhibits peptidyl transfer on both prokaryotic and eukaryotic ribosomes. The antibiotic inhibits the growth of Gram positive bacteria and various animal and insect cells. Fungi and Gram negative bacteria are resistant to Puromycin due to its low permeability. But in some particular conditions puromycin can be used for E. coli. For more than 30 years, puromycin has been widely used as a basic tool for studying protein synthesis. Now, puromycin hydrochloride is particularly useful for the selection of cell types harbouring plasmids carrying puromycin resistance genes.

Images & Validation

• Application Notes:
  Puromycin is usually used at concentrations ranging from 1 to 10 µg/mL. After transformation with a plasmid containing the pac gene, cells are incubated in their regular growth medium containing Puromycin to select for stable transfectants. 1) 48 hours post-transfection, pass cells (direct or diluted) in fresh medium containing Puromycin at the appropriate concentration. Note: Antibiotics work best when cells are actively dividing. If the cells become too dense, the antibiotic efficiency will decrease. It is best to split cells such that they are not more than 25% confluent. 2) Remove and replace antibiotic containing medium ever 3-4 days. 3) Evaluate cells for the formation of colonies after 7 days of selection. Colonies may require an additional week or more to develop depending on the host cell line and transfection/selection efficiency. 4) Transfer and pool 5-10 resistant clones to a 35mm cell culture plate and maintain on selection medium for an additional 7 days. This pooled culture will be expanded for subsequent cytotoxicity assays.

Storage & Handling

• Storage: Shipped at room temperature. Upon receipt it should be stored at 4°C. Puromycin is stable for up to three months at room temperature and at least one year at 4°C. For optimal stability and long term storage aqueous solutions can be stored at -20°C.
• Concentration: 10mg/ml
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only