FCCH100171 | FlowCellect™ Autophagy LC3 Antibody-based Assay Kit (100 tests)

FCCH100171
100 Tests  
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      Overview

      Replacement Information
      Description
      Catalogue NumberFCCH100171
      Trade Name
      • FlowCellect
      DescriptionFlowCellect™ Autophagy LC3 Antibody-based Assay Kit (100 tests)
      OverviewMillipore’s FlowCellect™ Autophagy LC3 Antibody-based Assay Kit provides a quantitative solution for the study of autophagy and potency evaluation of autophagy inducers using flow cytometry. This kit contains two key detection reagents to help facilitate the monitoring of lipidated LC3-II in a given cell system:

      1) The use of selective permeabilization solution discriminates between cytosolic LC3 from autophagic LC3 by extracting the soluble cytosolic proteins, while protecting LC3 which has been sequestered into the autophagosome;

      2) 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;

      By having the ability to measure and quantify autophagy, we are able to screen and rank order autophagy inducers or inhibitors, monitor cell culture health and protein turnover rate, study the mechanisms of protein degradation, and identity new autophagy targets and pathways leading to aging and neurodegenerative diseases.

      The anti-LC3 FITC conjugate and autophagy enabling reagents are optimized on guava® bench top flow cytometers and provide a complete solution for autophagy analysis. This kit can be used on any flow cytometer or imaging device by following the same protocol, offering researchers a reliable, quantitative, and fully validated solution to study autophagy. The anti-LC3 antibody provided in the kit has been carefully evaluated to ensure optimal performance, alleviating the need for any additional validation of the kit reagents.
      Alternate Names
      • FlowCellect
      Background InformationAutophagy is an intracellular catabolic pathway that causes cellular protein and organelle turnover, and it is associated in diverse diseases including Alzheimer's disease, aging, cancers, and Crohn's disease. It is a tightly regulated process that plays a normal part in cell growth, development, and cellular homeostasis. Autophagy functions as a housekeeping mechanism by disposing of aging and/or dysfunctional proteins and organelles through sequestering and priming proteins for lysosomal degradation. Increasing evidence suggests that not just apoptosis, but also autophagy, can contribute to cell death and greatly influence general cell health. Malfunctions of the autophagy process are proposed to influence cell health longevity and the capability of cells to function at full capacity.

      During autophagy, LC3 protein is translocated from the cytoplasm to the autophagosome where it is targeted to the lysosome for degradation. The process of autophagy can be categorized into four distinct stages (See figure 1):

      1) Induction and LC3 Translocation: The process is initiated by external/internal stimuli (e.g. nutrient depletion);
      2) Autophagosome formation: Unwanted cytosolic proteins and aging organelles are sequestered by a double membrane vesicle, i.e.—”autophagosome”. Formation of this vesicle is coordinated by complexes of Atg proteins (Atg5 and Atg12) that are conjugated, enabling the recruitment of LC3;
      3) Lysosomal docking and fusion: LC3 protein regulates traffic between autophagosome to lysosome. (LC3-I is cytoplasmic; LC3-II is lipidated and sequestered into autophagosomal membrane);
      4) Degradation: Fusion with the lysosome and subsequent breakdown of the autophagic vesicle and its contents.
      References
      Product Information
      Components
      • 1. 20X Anti-LC3 FITC, clone 4E12: (Part No. CS208214). One vial containing 600 L
      • 2. Autophagy reagent A: (Part No. CS208212) One vial (lyophilized)
      • 3. Autophagy reagent B: (Part No. CS208215) One vial containing 1 mL
      • 4. 5X Assay Buffer: (Part No. CS202124) One bottle containing 55 mL
      Detection methodFluorescent
      FormatCell Line
      Instrument compatibility
      • EasyCyte 8HT
      • EasyCyte
      Applications
      ApplicationThis FlowCellect Autophagy LC3 Antibody-based Assay Kit provides a quantitative solution for the study of autophagy & potency evaluation of autophagy inducers using flow cytometry.
      Key Applications
      • Flow Cytometry
      Biological Information
      Species Reactivity
      • Human
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      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
      Storage ConditionsAll reagents must be stored at 2 - 8ºC.

      All kit components are stable up to six (6) months from date of receipt if stored and handled correctly.
      Packaging Information
      Material Size100 Tests
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Brochure

      Title
      Advancing cancer research: From hallmarks & biomarkers to tumor microenvironment progression

      Technical Info

      Title
      Assessing Autophagy with the guava easyCyte™ Benchtop Flow Cytometer

      Posters

      Title
      Millipore Poster: Developing and optimizing a flow cytometry-based autophagy detection method for oncology drug development and biomarker identification

      User Guides

      Title
      FlowCellect™ Autophagy LC3 Antibody-based Assay Kit (100 tests)

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      Catalogue Number Description  
      0500-4120 InCyte Software for Guava® easyCyte HT Systems Show Pricing & Availability

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      Categories

      Life Science Research > Cell Analysis > guava easyCyte Flow Cytometers > Flow Cytometry Kits & Reagents