Anti-puromycin [3RH11]

Absolute Antibody

Anti-puromycin [3RH11]

7

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 (Frolinger et al., 2018; pmid: 29702026). 3RH11 offers advantageous alternative to traditional pulse-chase methods, which rely on radioactive amino acid labeling as it allows for the simple evaluation and quantification of translation directly using standard immunochemical methods (Balukoff et al., 2020; pmid: 33188200). 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 (Frolinger et al., 2018; pmid: 29702026). Practical comments: 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. 3RH11 is a very popular antibody used in countless experiments analysing global protein synthesis (mRNA translation) (Balukoff et al., 2020; pmid: 33188200; Crowell et al., 2016; pmid: 27464336; Khoutorsky et al., 2015; pmid: 26678009; Frolinger et al., 2018; pmid: 29702026; Wang et al., 2016; pmid: 27454289).

Functional Assay, ImmunoFluorescence, Western Blot, ELISA

Research Use Only

This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG1 format, for improved compatibility with existing reagents, assays and techniques.

-20°C,2°C to 8°C

12352203