This monoclonal antibody to puromycin provides a non-radioactive method to measure rates of global protein synthesis (mRNA translation) in cells or tissue slices incubated with puromycin, or animals treated with puromycin in vivo.
Puromycin is an aminonucleoside antibiotic, derived from the Streptomyces alboniger bacterium, that causes premature chain termination during translation taking place in the ribosome. Part of the molecule resembles the 3' end of the aminoacylated tRNA, making it useful for protein translation analysis.
Classical pulse-chase or flooding dose methods used to monitor protein synthesis rely on the measurement of radioactive methionine and cysteine labels. Analysis using puromycin immunodetection is an advantageous alternative to radioactive amino acid labeling, and allows for the evaluation/quantification of translation directly using standard immunochemical methods.
From the laboratory of Scot R. Kimball, PhD, Penn State College of Medicine
Read related blog post, Puromycin Incorporation as a Measure of Global Protein Synthesis »
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|Specificity:||This antibody recognizes puromycin.|
|Purity:||Protein G purified|
|Buffer:||PBS, 0.05% (w/v) Sodium azide|
|Tested Applications:||Western blotting (1:1,000), ELISA and Immunofluorescence microscopy.|
|Storage:||+4C (short-term), -20C (long-term); Avoid repeated freeze/thaw cycles.|
Scot R. Kimball, PhD
Penn State College of Medicine
Puromycin inhibits protein synthesis, which some cells may be more or less sensitive to. It is therefore highly recommended that the concentration of puromycin for protein translation assays be optimized for cell type. The suggested concentration of puromycin is 1uM, but for cells that are more sensitive, lower concentrations of puromycin may give better results.
Because cell growth (and protein synthesis) slows dramatically as cell near confluence, it is recommended that experiments are performed during growth phase (40-50% confluence). Label with an optimized concentration of puromycin for at least 20-30 min.
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