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ZMS1016

Sigma-Aldrich

Anti-Puromycin Antibody, clone 12D10 ZooMAb® Mouse Monoclonal

enhanced validationgreener alternative

recombinant, expressed in HEK 293 cells

biological source

mouse

Quality Level

recombinant

expressed in HEK 293 cells

conjugate

unconjugated

antibody form

purified antibody

antibody product type

primary antibodies

clone

12D10, recombinant monoclonal

description

recombinant, expressed in HEK 293 cells

product line

ZooMAb® learn more

form

lyophilized

mol wt

calculated mol wt Variable

species reactivity

human

species reactivity (predicted by homology)

all

packaging

antibody small pack of 25 μL

greener alternative product characteristics

Waste Prevention
Designing Safer Chemicals
Design for Energy Efficiency
Learn more about the Principles of Green Chemistry.

enhanced validation

recombinant expression
Learn more about Antibody Enhanced Validation

technique(s)

flow cytometry: suitable
immunocytochemistry: suitable
western blot: suitable

isotype

IgG2aκ

epitope sequence

Unknown

UniProt accession no.

greener alternative category

shipped in

ambient

storage temp.

2-8°C

General description

ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies.

Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them.

Learn more about ZooMAb here.

Specificity

Clone 12D10 is a ZooMAb® mouse recombinant monoclonal antibody that specifically detects puromycin incorporated into proteins.

Immunogen

Puromycin from Streptomyces alboniger.

Application

Quality Control Testing

Evaluated by Western Blotting in lysate from HEK293 cells treated with Puromycin.

Western Blotting Analysis (WB): A 1:50,000 dilution of this antibody detected Puromycin in lysate from HEK293 cells treated with Puromycin (25 μM, 10 min.), but not in cells treated with Puromycin (25 μM, 10 min) and Cycloheximide (25 μM, 10 min.).

Tested applications

Immunocytochemistry Analysis: 30 μg/mL from a representative lot detected Puromycin in HeLa treated with Puromycin.

Flow Cytometry Analysis: 0.1 μg from a representative lot detected Puromycin in HeLa cells treated with Puromycin.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Anti-Puromycin, clone 12D10 ZooMAb®, Cat. No. ZMS1016, is a recombinant Mouse monoclonal antibody that detects puromycin incorporated proteins and is tested for use in Flow Cytometry, Immunocytochemistry, and Western Blotting.

Target description

Puromycin is a naturally occurring aminonucleoside antibiotic that is derived from the Streptomyces alboniger bacterium, a gram-positive actinomycete. It acts as a protein synthesis inhibitor that blocks translation through premature chain termination. It attaches covalently to the C-terminus of nascent polypeptide chains, causing their premature termination and rapid dissociation from ribosomes. It is shown to act on both prokaryotic and eukaryotic cells. Its structure is similar to that of the 3 end of the aminoacyl-tRNA carrier of tyrosine or phenylalanine. Hence, it can occupy ribosomal site A and binds to the polypeptide chain synthesized by peptidyl transferase and bloc the entrance of the next aminoacyl-tRNA. Due to its non-selectivity and high systemic toxicity, puromycin is not used as a clinically relevant antibiotic, but is extensively used to study mechanisms of protein synthesis, particularly peptide bond formation and ribosome translocation. Monoclonal antibodies to puromycin can be used with standard immunochemical methods to directly monitor translation. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Aviner, R., et al. (2020). Comput. Struct. Biotechnol. J.18; 1074-1083).

Physical form

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL.

Reconstitution

30 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

Legal Information

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

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 Code

11 - Combustible Solids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

Enter Lot Number to search for Certificate of Origin (COO).

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

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