MABE343

Anti-Puromycin Antibody, clone 12D10

clone 12D10, from mouse

• Synonym(s): Anti-Puromycin, Clone 12D10 Anti-Puromycin, Puromycin Detection Antibody
• UNSPSC Code: 12352203
• NACRES: NA.41
• eCl@ss: 32160702
• Clone: 12D10, monoclonal
• Species reactivity: human
• Application: FACS, ICC, IF, IHC, IP, WB
• Citations: 657

Properties

• biological source: mouse
• Quality Segment: 100
• antibody form: purified immunoglobulin
• antibody product type: primary antibodies
• clone: 12D10, monoclonal
• species reactivity: human
• species reactivity (predicted by homology): all
• technique(s): flow cytometry: suitable, immunocytochemistry: suitable, immunofluorescence: suitable, immunohistochemistry: suitable, immunoprecipitation (IP): suitable, western blot: suitable
• isotype: IgG2aκ
• 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 weaker signal is observed in the co-presense of Cycloheximide, an inhibitor of protein biosynthesis in eukaryotic organisms.

Immunogen

Puromycin from Streptomyces alboniger

Application

Anti-Puromycin antibody, clone 12D10, detects puromycin incorporated into protein. Monoclonal antibodies to puromycin may be used with standard immunochemical methods.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
RNA Metabolism & Binding Proteins

Western Blotting Analysis (Total Protein Staining): HEK293 cell lysates treated with Puromycin and Cyclohexamide, or Puromycin only were resolved using SDS-PAGE and transferred to a membrane. Proteins were visualized using Ponceau S staining.

Immunocytochemistry Analysis: A 1:10,000 dilution from a representative lot detected Puromycin-incorporated neosynthesized proteins in HeLa cells treated with Puromycin.

Western Blotting Analysis: A representative lot detected Puromycin-incorporated neosynthesized proteins in WB (Reineke, L. C., et al. (2012). Mol Biol Cell. 23(18):3499-3510.; Trinh, M. A. et al. (2012). Cell Rep. 1(6):678-688.; Fortin, D. A., et al. (2012). J Neurosci. 32(24):8127-8137.; Clavarino, G., et al. (2012). PLoS Pathog. 8(5):e1002708.; David, A., et al. (2012). J Cell Biol. 197(1):45-57.; White, L. K., et al. (2011). J Virol. 85(1):606-620.; Hoeffer, C. A., et al. (2011). Proc Natl Acad Sci USA. 108(8):3383-3388.; Goodman, C. A., et al. (2010). FASEB J. 25(3):1028-1039.; Schmidt, E., K., et al. (2009). Nat Methods. 6(4):275-277.; Santini, E., et al. (2013). Nature. 493(7432):411-415.; Quy. P. N., et al. (2013). J Biol Chem. 288(2):1125-1134.; Hulmi. J. J., et al. (2012). Am J Physiol Endocrinol Metab. 304(1):E41-50.; Bhattacharya, A., et al. (2012). Neuron. 76(2):325-337.; Hoeffer, C. A., et al. (2013). J Neurophysiol. 109(1):68-76.).

Immunofluorescence Analysis: A representative lot detected Puromycin-incorporated neosynthesized proteins in WB (Reineke, L. C., et al. (2012). Mol Biol Cell. 23(18):3499-3510.; Trinh, M. A. et al. (2012). Cell Rep. 1(6):678-688.; Fortin, D. A., et al. (2012). J Neurosci. 32(24):8127-8137.;David, A., et al. (2012). J Cell Biol. 197(1):45-57.; David, A., et al. (2011). J Biol Chem. 286(23):20688-20700.; White, L. K., et al. (2011). J Virol. 85(1):606-620.; Hoeffer, C. A., et al. (2011). Proc Natl Acad Sci USA. 108(8):3383-3388.; Schmidt, E., K., et al. (2009). Nat Methods. 6(4):275-277.; Goodman, C. A., et al. (2012). Proc Natl Acad Sci USA. 109(17):E989.; Santini, E., et al. (2013). Nature. 493(7432):411-415.; Quy. P. N., et al. (2013). J Biol Chem. 288(2):1125-1134.).

Immunohistochemistry Analysis: A representative lot detecte Puromycin-incorporated neosynthesized protein in IHC (Goodman, C. A., et al. (2010). FASEB J. 25(3):1028-1039.).

Fluorescence Activated Cell Sorting Analysis: A representative lot 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

Anti-Puromycin Antibody, clone 12D10 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

Protein G Purified

Purified mouse monoclonal IgG2aκ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Preparation Note

Stable for 1 year at 2-8°C from date of receipt.

Analysis Note

Control
HEK293 cell lysates treated with Puromycin and Cyclohexamide, or with Puromycin only.

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

Western Blotting Analysis: A 1:25,000 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-incorporated neosynthesized proteins in HEK293 cells treated with Puromycin and Cyclohexamide.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

ALEXA FLUOR is a trademark of Life Technologies

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Safety Information

• Storage Class: 12 - Non Combustible Liquids
• wgk: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable