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Purified anti-Puromycin Antibody

Pricing & Availability
Clone
2A4 (See other available formats)
Regulatory Status
RUO
Other Names
Puromycin from streptomyces alboniger
Isotype
Mouse IgG2a, κ
Ave. Rating
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Product Citations
publications
A. 2A4_PURE_Puromycin_Antibody_041222
Human peripheral blood mononuclear cells were stimulated with Cell Activation Cocktail (Cat. No. 423301) in the presence of Puromycin (1 µg/mL) (filled dark purple histogram) or without (open histogram) for 4hours or left unstimulated (light purple histogram), treated with True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401) and then stained with purified anti-Puromycin (clone 2A4) followed by PE anti-mouse IgG.
  • A. 2A4_PURE_Puromycin_Antibody_041222
    Human peripheral blood mononuclear cells were stimulated with Cell Activation Cocktail (Cat. No. 423301) in the presence of Puromycin (1 µg/mL) (filled dark purple histogram) or without (open histogram) for 4hours or left unstimulated (light purple histogram), treated with True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401) and then stained with purified anti-Puromycin (clone 2A4) followed by PE anti-mouse IgG.
  • B. 2A4_PURE_Puromycin_Antibody_2_041222
    HeLa cells were treated with Puromycin (10 µg/mL) (left) for 2 hours or left untreated (right) and fixed with 4% paraformaldehyde 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) overnight at 4°C followed by Alexa Fluor® 555 goat anti-mouse IgG (red) for one hour at room temperature. Nuclei were counterstained with DAPI (blue)
  • C. 381502-figC
    Whole cell extracts (15 µg total protein) prepared from HeLa cells treated with 1 µg/mL Puromycin (+Puro) or vehicle (-Puro) for 1 hour or were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 1 µg/mL (1:500 dilution) of Purified anti-Puromycin antibody (clone 2A4) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin antibody (Cat. No. 607903) was used as a loading control at a 1:25000 dilution (lower). Western-Ready™ ECL Substrate Plus Kit (Cat. No. 426317) was used as a detection agent. Lane M: Molecular weight marker
  • D. 381502-figD
    Whole cell extracts (15 µg total protein) prepared from HeLa cells treated with 1 µg/mL Puromycin (+Puro) or vehicle (-Puro) for 1 hour or were resolved on a 4-12% Bis-Tris gel, transferred to a PVDF membrane, and probed with 1 µg/mL (1:500 dilution) of Purified anti-Puromycin antibody (clone 2A4) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-β-actin antibody (Cat. No. 607903) was used as a loading control at a 1:25000 dilution (lower). Western-Ready™ ECL Substrate Plus Kit (Cat. No. 426317) was used as a detection agent. Lane M: Molecular weight marker
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381502 100 µg 429 CHF
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Description

Puromycin is an aminonucleoside antibiotic, derived from the Streptomyces alboniger bacterium, which functions as a Tyr-tRNA mimetic that enters the ribosome A site and incorporates into the nascent chain C terminus. Antibodies against Puromycin can detect puromycylated nascent chains released from ribosomes and thereby quantify protein translation. This monoclonal antibody to puromycin can be used in flow cytometry, immunofluorescence or immunoblotting to measure rates of global protein synthesis (mRNA translation) in cells incubated with Puromycin. 

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse, Rat, All species
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Puromycin from Streptomyces alboniger coupled to KLH
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

ICFC - Quality tested
ICC, WB, IP - Verified
Recommended Usage

Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤ 0.125 µg per million cells in 100 µL volume. For immunocytochemistry, a concentration of 2.5 μg/mL is recommended. For western blotting, the suggested use of this reagent is 1.0 µg/mL. For immunoprecipitation, the suggested use of this reagent is 2.0 µg/test. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

It is recommended that the timing and concentration of Puromycin treatment be optimized for each cell type. 1.0 - 10.0 µg/mL of Puromycin for 30 minutes to 2 hours are suggested ranges for activated PBMCs or cell lines in growth phase.

For flow cytometry, the amount of anti-puromycin antibody may also have to be optimized. Cells with high levels of protein translation require lower concentrations of antibody.  

RRID
AB_2924575 (BioLegend Cat. No. 381502)

Antigen Details

Distribution

Cytoplasm, Ribosomes

Cell Type
Embryonic Stem Cells
Biology Area
Cell Biology, Protein Synthesis
Antigen References
  1. Schmidt E, et al. 2009. Nat Methods. 6:275-7.
  2. Arguello R, et al. 2020. Cell Metabolism. 32:1063-1075.
Gene ID
NA
UniProt
View information about Puromycin on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 04.12.2022

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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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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