MilliporeSigma
All Photos(5)

Documents

MABE343

Sigma-Aldrich

Anti-Puromycin Antibody, clone 12D10

clone 12D10, from mouse

Sign Into View Organizational & Contract Pricing

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)

Related Categories

Compare Similar Items

View Full Comparison

Show Differences

1 of 4

This Item
MABE341AB3258MAB2247
antibody form

purified immunoglobulin

antibody form

purified antibody

antibody form

purified immunoglobulin

antibody form

purified immunoglobulin

biological source

mouse

biological source

rat

biological source

goat

biological source

mouse

species reactivity

human

species reactivity

human

species reactivity

rat, mouse

species reactivity

human

clone

12D10, monoclonal

clone

17H1, monoclonal

clone

polyclonal

clone

12G10, monoclonal

technique(s)

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

technique(s)

western blot: suitable

technique(s)

immunocytochemistry: suitable, western blot: suitable

technique(s)

ELISA: suitable, cell culture | mammalian: suitable, flow cytometry: suitable, immunoprecipitation (IP): suitable, western blot: suitable

Still not finding the right product?  

Give our Product Selector Tool a try.

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

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.

Storage Class

12 - Non Combustible Liquids

wgk_germany

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’.

Already Own This Product?

Documents related to the products that you have purchased in the past have been gathered in the Document Library for your convenience.

Visit the Document Library

Difficulty Finding Your Product Or Lot/Batch Number?

Product numbers are combined with Pack Sizes/Quantity when displayed on the website (example: T1503-25G). Please make sure you enter ONLY the product number in the Product Number field (example: T1503).

Example:

T1503
Product Number
-
25G
Pack Size/Quantity

Additional examples:

705578-5MG-PW

PL860-CGA/SHF-1EA

MMYOMAG-74K-13

1000309185

enter as 1.000309185)

Having trouble? Feel free to contact Technical Service for assistance.

Lot and Batch Numbers can be found on a product's label following the words 'Lot' or 'Batch'.

Aldrich Products

  • For a lot number such as TO09019TO, enter it as 09019TO (without the first two letters 'TO').

  • For a lot number with a filling-code such as 05427ES-021, enter it as 05427ES (without the filling-code '-021').

  • For a lot number with a filling-code such as STBB0728K9, enter it as STBB0728 without the filling-code 'K9'.

Not Finding What You Are Looking For?

In some cases, a COA may not be available online. If your search was unable to find the COA you can request one.

Request COA

Customers Also Viewed

Slide 1 of 2

1 of 2

Thomas Chaillou et al.
Journal of cellular physiology, 231(9), 1894-1902 (2015-12-20)
The ribosome has historically been considered to have no cell-specific function but rather serve in a "housekeeping" capacity. This view is being challenged by evidence showing that heterogeneity in the protein composition of the ribosome can lead to the functional
Preeti Shahi et al.
PLoS neglected tropical diseases, 10(1), e0004340-e0004340 (2016-01-07)
Entamoeba histolytica is an obligate protozoan parasite of humans, and amebiasis, an infectious disease which targets the intestine and/or liver, is the second most common cause of human death due to a protozoan after malaria. Although amebiasis is usually asymptomatic
Hanna Hörnberg et al.
The Journal of neuroscience : the official journal of the Society for Neuroscience, 36(50), 12697-12706 (2016-12-16)
The establishment of precise topographic maps during neural development is facilitated by the presorting of axons in the pathway before they reach their targets. In the vertebrate visual system, such topography is seen clearly in the optic tract (OT) and
Daniel W D West et al.
The Journal of physiology, 594(2), 453-468 (2015-11-10)
Ribosome biogenesis is the primary determinant of translational capacity, but its regulation in skeletal muscle following acute resistance exercise is poorly understood. Resistance exercise increases muscle protein synthesis acutely, and muscle mass with training, but the role of translational capacity
Anna Maria Copeland et al.
Journal of virology, 87(21), 11659-11669 (2013-08-24)
Rift Valley fever virus (RVFV), an ambisense member of the family Bunyaviridae, genus Phlebovirus, is the causative agent of Rift Valley fever, an important zoonotic infection in Africa and the Middle East. Phlebovirus proteins are translated from virally transcribed mRNAs

Articles

Leverage the inherent immune ability of an animal to generate antibodies that bind to specific molecules.

Explore the basics of working with antibodies including technical information on structure, classes, and normal immunoglobulin ranges.

Uses for immunofluorescence (IF)—where an antibody conjugated to a molecule that fluoresces when excited by lasers— include protein localization, confirmation of post-translational modification or activation, and proximity to/complexing with other proteins.

Antibodies combine with specific antigens to generate an exclusive antibody-antigen complex. Learn about the nature of this bond and its use as a molecular tag for research.

See All

Protocols

Tips and troubleshooting for FFPE and frozen tissue immunohistochemistry (IHC) protocols using both brightfield analysis of chromogenic detection and fluorescent microscopy.

Learn key steps in flow cytometry protocols to make your next flow cytometry experiment run with ease.

Explore our flow cytometry guide to uncover flow cytometry basics, traditional flow cytometer components, key flow cytometry protocol steps, and proper controls.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service