How do you dilute puromycin?

How do you dilute puromycin?

If necessary, prepare an aliquot of 1mg/ml puromycin (working concentration) by diluting a 10mg/ml stock 1:10 with sterile water.

How do I select cells with puromycin?

Prior to using the puromycin antibiotic, titrate the selection agent to determine the optimal concentration for target cell line. Use the lowest concentration that kills 100% of non-transfected cells in 3-5 days from the start of puromycin selection.

What is mechanism of action of puromycin?

Puromycin is a naturally occurring aminonucleoside antibiotic that inhibits protein synthesis by ribosome-catalyzed incorporation into the C-terminus of elongating nascent chains, blocking further extension and resulting in premature termination of translation.

How does streptomycin inhibit protein synthesis?

Streptomycin irreversibly binds to the 16S rRNA and S12 protein within the bacterial 30S ribosomal subunit. As a result, this agent interferes with the assembly of initiation complex between mRNA and the bacterial ribosome, thereby inhibiting the initiation of protein synthesis.

Is puromycin soluble in water?

The product is soluble in water (50 mg/ml), yielding a clear, colorless to faint yellow solution. The stock solution may be passed through a 0.22 µm filter and stored in aliquots at –20 °C. It is also soluble in methanol (10 mg/ml).

How puromycin inhibits protein synthesis?

Is puromycin light sensitive?

Puromycin Dihydrochloride is light sensitive on par with the light sensitivity of most basal media like DMEM and RPMI 1640. We would recommend limiting exposure of this product to light as much as possible (i.e,. don’t leave on the bench or under hood lights longer than necessary).

What is the puromycin resistance gene?

Since puromycin is toxic to the growth of various eukaryote cells including mammalian cells, a puromycin-resistant gene (puromycin-N-acetyltransferase gene (pac)) is commonly used as a selection marker gene in mammalian cells (de la Luna et al., 1988, Vara et al., 1986).

What is the molecular weight of puromycin?

544.43
Technical Data