General description
We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency".
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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.