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Puromycin dihydrochloride

Pricing & Availability
Regulatory Status
RUO
Other Names
Protein Synthesis Inhibitor, Puromycin 2HCl, C22H31Cl2N7O5
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Product Citations
publications
a. Puromycin-dihydrochloride_1_091724
Flow Cytometric Analysis of Intracellular Puromycin in Cells Treated with Inhibitors. Human PBMCs were stimulated with immobilized Ultra-LEAF™ Purified anti-human CD3 (clone UCHT1) (Cat. No. 300437) and Ultra-LEAF™ purified CD28 (clone CD28.2) (Cat. No. 302933) for 24 hours before being treated with 10 mM 2-Deoxy-D-Glucose (2-DG), 1 µM Oligomycin (O) or a combination of both (DGO) for 30 minutes at 37°C. Control cells were left untreated while test samples were treated with Puromycin (10 µg/mL) for 15 minutes. Cells were pelleted and resuspended in Cyto-Fast™ Fix/Perm Solution (Cat. No. 426803). After 30 minutes, cells were washed in Cyto-Fast™ Perm Wash Solution and intracellularly stained with PE anti-Puromycin (Cat. No. 381503). PE-fluorescence was analyzed by flow cytometry and the relative fluorescence was calculated by normalizing the mean fluorescence intensity of inhibitor-treated cells to vehicle-treated cells (Control). Error bars represent standard deviation from 3 different donors.
  • a. Puromycin-dihydrochloride_1_091724
    Flow Cytometric Analysis of Intracellular Puromycin in Cells Treated with Inhibitors. Human PBMCs were stimulated with immobilized Ultra-LEAF™ Purified anti-human CD3 (clone UCHT1) (Cat. No. 300437) and Ultra-LEAF™ purified CD28 (clone CD28.2) (Cat. No. 302933) for 24 hours before being treated with 10 mM 2-Deoxy-D-Glucose (2-DG), 1 µM Oligomycin (O) or a combination of both (DGO) for 30 minutes at 37°C. Control cells were left untreated while test samples were treated with Puromycin (10 µg/mL) for 15 minutes. Cells were pelleted and resuspended in Cyto-Fast™ Fix/Perm Solution (Cat. No. 426803). After 30 minutes, cells were washed in Cyto-Fast™ Perm Wash Solution and intracellularly stained with PE anti-Puromycin (Cat. No. 381503). PE-fluorescence was analyzed by flow cytometry and the relative fluorescence was calculated by normalizing the mean fluorescence intensity of inhibitor-treated cells to vehicle-treated cells (Control). Error bars represent standard deviation from 3 different donors.
  • b. Puromycin-dihydrochloride_2_091724
    HeLa cells were treated with Puromycin (10 µg/mL) for 2 hours and fixed with Fixation Buffer (Cat. No. 420801) for 30 minutes, permeabilized with 0.5% Triton X-100 for 15 minutes, and blocked with 5% FBS for 30 minutes. Cells were then intracellularly stained with 2.5 µg/mL Purified anti-Puromycin (clone 2A4) (Cat. No. 381502) overnight at 4°C followed by Alexa Fluor® 555 Goat anti-mouse IgG (red) (Cat. No. 405324) for one hour at room temperature. Nuclei were counterstained with DAPI (blue) (Cat. No. 422801). Scale bar: 20 µm
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427508 50 mg 120€
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Description

Puromycin dihydrochloride is a naturally occurring amino nucleoside antibiotic produced by Streptomyces alboniger. Comprised of a nucleoside covalently bound to an amino acid, Puromycin mimics the 3′ end of aminoacylated tRNAs and thus inhibits protein synthesis by ribosome-catalyzed incorporation into elongating nascent chains, which results in premature termination of translation. It is used as a selection agent in mammalian cell cultures for cell lines that express a puromycin resistance via puromycin N-acetyl-transferase (pac).

Puromycin immunodetection with antibodies such as BioLegend’s clone 2A4 allows the direct evaluation of translational activity in single cells by immunofluorescence microscopy and flow cytometry.

Product Details
Technical data sheet

Product Details

Formulation
Lyophilized
Preparation
Solubility: 50-100 mg/mL in distilled deionized water, PBS or DMSO.
Storage & Handling
Store lyophilized powder at -20°C. When reconstituted, aliquot and store at -20°C. Use reconstituted material within 6 months to avoid loss of potency.
Application

Cell Culture - Verified
Recommended Usage

Working concentrations of Puromycin dihydrochloride for cell-based assays range from 0.2 – 10 μg/mL. It is recommended that the reagent be titrated for each experiment to determine optimal performance.

Antigen Details

Structure
C22 H29N7O5 · 2HCl. MW = 544.43 g/mol
Function
Protein synthesis inhibitor, antibiotic
Antigen References
  1. Arguello R, et al. 2020. Cell Metabolism. 32:1063-1075.
  2. Nathans D, et al. 1964. PNAS.51:585-92.
  3. Miyamoto-Sato E, et al. 2000. Nucleic Acids Res. 28:1176-82.  
  4. Schmidt EK, et al. 2009. Nat Methods. 6:275-7.
Gene ID
NA

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Go To Top Version: 1    Revision Date: 09.17.2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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