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Key Documents

MABE343

Sigma-Aldrich

Anti-Puromycin Antibody, clone 12D10

clone 12D10, from mouse

Synonim(y):

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

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About This Item

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

pochodzenie biologiczne

mouse

Poziom jakości

forma przeciwciała

purified immunoglobulin

rodzaj przeciwciała

primary antibodies

klon

12D10, monoclonal

reaktywność gatunkowa

human

reaktywność gatunkowa (przewidywana na podstawie homologii)

all

metody

flow cytometry: suitable
immunocytochemistry: suitable
immunofluorescence: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

izotyp

IgG2aκ

Warunki transportu

wet ice

docelowa modyfikacja potranslacyjna

unmodified

informacje o genach

human ... NPEPPS(9520)

Opis ogólny

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.

Specyficzność

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

Zastosowanie

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.

Jakość

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.

Opis wartości docelowych

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.

Postać fizyczna

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.

Przechowywanie i stabilność

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

Komentarz do analizy

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

Inne uwagi

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

Informacje prawne

ALEXA FLUOR is a trademark of Life Technologies

Oświadczenie o zrzeczeniu się odpowiedzialności

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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Kod klasy składowania

12 - Non Combustible Liquids

Klasa zagrożenia wodnego (WGK)

WGK 1

Temperatura zapłonu (°F)

Not applicable

Temperatura zapłonu (°C)

Not applicable


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