MABE342

Anti-Puromycin Antibody, clone 4G11

clone 4G11, from mouse

• UNSPSC Code: 12352203
• NACRES: NA.41
• eCl@ss: 32160702
• Clone: 4G11, monoclonal
• Species reactivity: human
• Application: FACS, ICC, IF, WB
• Citations: 21

Properties

• biological source: mouse
• Quality Segment: 100
• antibody form: purified immunoglobulin
• antibody product type: primary antibodies
• clone: 4G11, monoclonal
• species reactivity: human
• species reactivity (predicted by homology): all
• technique(s): flow cytometry: suitable, immunocytochemistry: suitable, immunofluorescence: suitable, western blot: suitable
• isotype: IgG1κ
• shipped in: wet ice
• target post-translational modification: unmodified
• Gene Information: human ... NPEPPS(9520)

Description

General description

Puromycin is an aminonucleoside antibiotic, derived from the Streptomyces alboniger bacterium, that functions as a protein synthesis inhibitor that blocks translation through premature chain termination in the ribosome. Monoclonal antibodies to puromycin may be used with standard immunochemical methods to directly monitor translation, a method known as surface sensing of translation (SUnSET). Part of the molecule resembles the 3′ end of the aminoacylated tRNA, making it useful for protein translation analysis. Puromycin induces DNA fragmentation in thymocytes and in human HL-60 leukemia cells.

Puromycin is incorporated in neosynthesized proteins. In the presense of Puromycin only, this antibody detects Puromycin-incorporated neosynthesized proteins at multiple molecular weights. However, a decrease in signal is observed in the co-presense of Cycloheximide, an inhibitor of protein biosynthesis in eukaryotic organisms.

Application

Anti-Puromycin antibody, clone 4G11 detects puromycin incorporated into protein. Monoclonal antibodies to puromycin may also be used with standard immunochemical methods.

Immunocytochemistry Analysis: A 1:2,000 dilution from a representative lot detected Puromycin-incorporated neosynthesized proteins in HeLa cells treated with Puromycin. However, a decrease in signal is observed with the addition of Cycloheximide, an inhibitor of protein biosynthesis in eukaryotic organisms.

Western Blotting Analysis: A representative lot from an independent laboratory detected Puromycin-incorporated neosynthesized proteins in WB (David, A., et al. (2012). J Cell Biol. 197(1):45-57.; Schmidt, E., K., et al. (2009). Nat Methods. 6(4):275-277.).

Immunofluorescence Analysis: A representative lot from an independent laboratory detected Puromycin-incorporated neosynthesized proteins in IF (David, A., et al. (2012). J Cell Biol. 197(1):45-57.; Schmidt, E., K., et al. (2009). Nat Methods. 6(4):275-277.).

Fluorescence Activated Cell Sorting Analysis: A representative lot from an independent laboratory detected Puromycin-incorporated neosynthesized proteins in FACS (David, A., et al. (2012). J Cell Biol. 197(1):45-57.; Schmidt, E., K., et al. (2009). Nat Methods. 6(4):275-277.).

Alexa Fluor^™ is a registered trademark of Life Technologies.

Biochem/physiol Actions

Demonstrated to react with Human test sample, preincubated with Puromycin. Predicted to react with all species when test sample is incubated with Puromycin.

Physical form

Format: Purified

Analysis Note

Evaluated by Western Blotting in HEK293 cell lysates treated with Puromycin and Cyclohexamide, or with Puromycin only.

Western Blotting Analysis: A 1:12,500 dilution of this antibody detected Puromycin-incorporated neosynthesized proteins in HEK293 cell lysates treated with Puromycin only. This antibody also detected small mounts of puromycin in HEK293 cells treated with Puromycin and Cyclohexamide.

Legal Information

ALEXA FLUOR is a trademark of Life Technologies

Safety Information

• Storage Class: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point (°F): Not applicable
• Flash Point (°C): Not applicable