MCH200110 Muse™RFP-LC3 Reporter Autophagy Assay Kit

MCH200110
100 Tests  
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      Overview

      Replacement Information
      Description
      Catalogue NumberMCH200110
      Trade Name
      • Muse
      DescriptionMuse™RFP-LC3 Reporter Autophagy Assay Kit
      OverviewEMD Millipore’s Muse™ RFP-LC3 Reporter Autophagy Assay kit provides a quantitative solution for the study of autophagy and potency evaluation of autophagy inducers using flow cytometry. This kit has four unique features:
      1)The use of selective permeabilization solution discriminates cytosolic LC3 from autophagic LC3 by extracting the soluble cytosolic proteins, while protecting LC3 which has been sequestered into the autophagosome from degradation;
      2) A monomeric RFP is used as a reporter to facilitate the translocation of the fusion protein. Other forms of RFP are not preferred as they form dimers and aggregate when over-expressed in the cells, preventing their extraction from the cytoplasm and impairing the ability to measure translocation by flow cytometry;
      3) Since autophagy is a constitutive cellular degradation process, the use of an autophagy detection reagent (Autophagy Reagent A) will prevent the lysosomal degradation of LC3, allowing its quantification by flow cytometry;
      4) The monomeric RFP used in our fusion protein to LC3 is attached on the 5' end (N-terminal fusion), protecting the RFP from Atg4 cleavage, thus, allowing its visualization within the autophagosomes

      To assist in autophagy study, U20S (human osteosarcoma) cells are stably transfected, which are suitable for quantitative and higher throughput analysis of autophagy. Data generated using the Muse™ Cell Analyzer along with the corresponding Muse™ software module provides statistical values measuring:

      • Mean Autophagy Value (for both control and test samples)
      • Autophagy Induction Ratio (Test sample fluorescence relative to control)
      • Percentage of cells with increased autophagy (Test sample versus control)

      EMD Millipore’s Muse™ RFP-LC3 Reporter Autophagy Assay kit includes one immortalized RFP-LC3 reporter cell line to measure and track the levels of LC3 within the cell. The RFP-LC3 reporter cell line provided in the kit has been carefully optimized along with the corresponding reagents to ensure optimal performance. This kit contains reagents A and B, along with an assay buffer to provide researchers a complete solution for autophagy analysis. The autophagy detection reagents and cell line have been optimized together to ensure the ability to measure and discriminate between cytosolic and lipidated LC3 to accurately measure the autophagic process. Sufficient enabling reagents are provided to perform 100 tests
      The Muse™ RFP-LC3 Reporter Autophagy Assay kit is optimized on the Muse™ Cell Analyzer. The Muse™ System makes sophisticated fluorescent-based analysis fast, easy, convenient, and affordable. Sample preparation is minimal, and after loading samples onto the Muse™ Cell Analyzer, the intuitive software provides detailed or summary analysis of your cell sample in a few short steps.
      For Research Use Only; not for use in diagnostic procedures.
      References
      Product Information
      Instrument compatibility
      • Muse Cell Analyzer
      Applications
      ApplicationThe Muse® RFP-LC3 Reporter Autophagy Assay kit includes one immortalized RFP-LC3 reporter cell line to measure and track the levels of LC3 within the cell. The RFP-LC3 reporter cell line provided in the kit has been optimized along with the corresponding reagents to ensure optimal performance.
      Biological Information
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      Materials Information
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      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • 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 and Shipping Information
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      Material Size100 Tests
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      Documentation

      Brochure

      Title
      Muse Cell Analyzer Service Plans (MilliporeSigma)
      Experience simple, affordable flow cytometry.

      Technical Info

      Title
      White Paper: Individual cell analysis for protein expression and cell health markers presents a rapid, quantitative alternative to bulk detection methods for fundamental assays.

      Posters

      Title
      Muse Cell Health Poster: A World of Cell Health Between Life and Death (MilliporeSigma)

      User Guides

      Title
      Muse™ RFP-LC3 Reporter Autophagy Assay Kit User’s Guide