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MABE343

Sigma-Aldrich

Anti-Puromycin Antibody, clone 12D10

clone 12D10, from mouse

Synonym(s):

Anti-Puromycin, Clone 12D10 Anti-Puromycin, Puromycin Detection Antibody

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

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)

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.

Specificity

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.

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.

Quality

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.

Target description

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.

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.

Storage and Stability

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.

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.

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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