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  • Who we serve
    • Academia
      Explore BioLegend

      Learn about our world class antibodies for a diverse set of research areas including immunology, neuroscience, cancer, stem cells and cell biology.

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    • Pharma / Biotech
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    • Clinical Laboratories
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    • Healthcare Professionals
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    • Contract Research Organizations
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  • Products
    • Research & Development
      • Genomic Analysis
        • IncRNA in Functional Studies
        • Nucleic Acid Isolation
        • NGS Workflows
        • Microplate Readers
        • Microfluidic Nucleic Acid Analysis
        • CRISPR Technologies
        • RNA Interference
        • Transfection and Ancillary Reagents
        • Oligonucleotide Custom Synthesis
        • cDNAs and ORF Clones
        • Single-Cell Sequencing
        • Labeled Nucleotides
        • qPCR
        • Library Prep Kits
        • Magnetic Beads
        • Viral Vector Products
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      • Protein Analysis
        • Binding Assays
        • Immunoassays
        • Microfluidic Protein Characterization
        • Microplate Readers
        • Microplates
        • Recombinant Proteins
        • Western Blotting
        • Sample Dissociation
        • M-PVA Magnetic Beads
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      • Cell Analysis
        • Cell Isolation
        • Cell Lines & Stem Cells
        • Cell Counting and Image Cytometry
        • Cell Health & Viability
        • Cellular Imaging & Analysis
        • Immunoassays
        • In Vivo Imaging
        • Microplates
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      • Research Solutions
        • Immunoassays Solutions
        • pHSense Reagents
        • Biomarker Discovery
        • Cell and Gene Therapy
        • GMP Workflows
        • Biologics
        • Small Molecule Drug Discovery
        • Disease Research
        • Target Class
        • Drug Development
        • Precision Medicine Research
        • Functional Genomic Screening Solutions
        • Assay Development Workflows
        • Physiological Model Solutions
        • Biobanking Workflows
        • Agrigenomics Workflows
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    • Clinical & Diagnostics
      • Reproductive Health
        • Maternal and Prenatal Testing
        • Newborn Screening
        • Newborn Sequencing Research
        • Pregnancy-Relevant Infections
        • Endocrine Reproductive
        • Cell-free DNA Analysis
        • Neonatal Research
        • Preimplantation Genetic Testing
        • PlGF Testing Research
        • Molecular Cytogenetics
        New IVD Mimix™ reference standards for oncology workflows.

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      • Infectious Diseases
        • Tuberculosis Management
        • EUROIMMUN Solutions
        • IDS Solutions
        • Nucleic Acid Isolation for Pathogen Detection
        • Cytomegalovirus
        • SARS-COV-2 Testing Solutions
        • Bacterial & Viral Nucleic Acid Isolation
        • Metagenomics
        New IVD Mimix™ reference standards for oncology workflows.

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      • Cancer
        • ASR Flow Cytometry Antibodies
        • HPV Testing
        • ctDNA Workflows
        • miRNA-seq Analysis
        • Exosome/cfRNA Analysis
        • Targeted Sequencing
        • Mimix Reference Standards
        • Functional Testing
        NEW! Mimix IVD reference standards

        Providing diagnostic labs with trusted quality controls for developing tests and monitoring workflows, with the added assurance of IVD marking. Explore our newest MimixTM controls.

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      • Autoimmunity
        • EUROIMMUN Solutions
        • IDS Solutions
        New IVD Mimix™ reference standards for oncology workflows.

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      • Endocrinology
        New IVD Mimix™ reference standards for oncology workflows.

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      • Allergy
        New IVD Mimix™ reference standards for oncology workflows.

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      • Neurodegeneration
        • Alzheimer's Disease
        • Amytrophic Lateral Sclerosis (ALS)
        • Neurogranin
        • BACE1
        New IVD Mimix™ reference standards for oncology workflows.

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      • Rapid Patient Testing
        New IVD Mimix™ reference standards for oncology workflows.

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      New IVD Mimix™ reference standards for oncology workflows.

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    • Reagents
      • BioLegend
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      • Cellular Imaging Reagents
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      • chemagic Nucleic Acid Purification Kits
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      • CRISPR Technologies
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      • GPCR Research Reagents
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      • Immunoassays
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      • In Vivo Imaging Reagents
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      • Liquid Scintillation Cocktails
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      • NGS Library Prep Kits
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      • pHSense Reagents
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      • qPCR
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      • Radiochemicals
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      • RNA Interference
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      • Primary and Secondary Antibodies
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    • Platforms & Automation
      • Nucleic Acid Purification
        • chemagic Applications
        • chemagic IVD Instruments
        • chemagic IVD Kits
        • chemagic Instruments
        • chemagic Kits
        • M-PVA Magnetic Beads
        New! Cellometer™ Ascend™

        Explore our cell counting and image cytometry solutions.

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      • Automated Liquid Handling
        • Liquid Handling Workstations
        • Automated Liquid Handling Applications
        • Liquid Handling Consumables & Accessories
        New! Cellometer™ Ascend™

        Explore our cell counting and image cytometry solutions.

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      • Integrated Lab Automation
        • Automated Plate Loading
        • Workstation Modules
        • Scheduling & Control Software
        • Robotics
        New! Cellometer™ Ascend™

        Explore our cell counting and image cytometry solutions.

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      • Microfluidic Analysis
        • Microfluidic Protein Characterization
        • Microfluidic Nucleic Acid Analysis
        New! Cellometer™ Ascend™

        Explore our cell counting and image cytometry solutions.

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      • Detection Solutions
        • Plate Reader Accessories
        • Microplate Readers
        • Radiometric Detectors
        • Radiochemicals
        • Liquid Scintillation Cocktails
        • Radiometric Consumables & Accessories
        New! Cellometer™ Ascend™

        Explore our cell counting and image cytometry solutions.

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      • Imaging
        • Cellular Imaging & Analysis
        • In Vivo Imaging
        New! Cellometer™ Ascend™

        Explore our cell counting and image cytometry solutions.

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      • Cell Counting and Image Cytometry
        New! Cellometer™ Ascend™

        Explore our cell counting and image cytometry solutions.

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      • Sample Homogenization
        • Homogenizers
        • Homogenizer Accessories
        • Homogenizer Consumables
        • Sample Centrifugation
        • Sample Sonication
        New! Cellometer™ Ascend™

        Explore our cell counting and image cytometry solutions.

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      • In Vitro Diagnostics (IVD) Platforms & Automation
        • IVD Nucleic Acid Isolation
        • IVD Automated Liquid Handling
        • IVD Radiometric Detectors
        • EUROIMMUN Instruments
        • IDS Instruments
        • Clinical Applications
        New! Cellometer™ Ascend™

        Explore our cell counting and image cytometry solutions.

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      • Consumables & Accessories
        • Cassettes
        • Cell Harvesters
        • In Vivo Imaging Accessories
        • High-Content Imaging Accessories
        • Microplates
        • Radioactive Spill Cleaners
        • Scintillation Cocktails
        • Transfer Membranes
        • Charcoal Traps
        • Homogenizer Accessories
        • Homogenizer Consumables
        New! Cellometer™ Ascend™

        Explore our cell counting and image cytometry solutions.

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      • Instrument Service & Maintenance
        • On Demand Equipment Service
        • Equipment Service Plans
        New! Cellometer™ Ascend™

        Explore our cell counting and image cytometry solutions.

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      New! Cellometer™ Ascend™

      Explore our cell counting and image cytometry solutions.

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    • Consumables & Accessories
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    • Signals Software
      • All Products
        • Signals One
        • ChemDraw
        • Spotfire®
        • Signals Clinical
        • Signals Notebook
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      • All Solutions
        • Biology
        • Discovery Chemistry
        • Clinical & Translational
        • Specialty Chemicals
        • Food/Flavor/Fragrance
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    • Revvity Omics Services
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  • Services
    • Preclinical Services
      • Antibody Drug Conjugate Services
        Preclinical services

        Work with our experienced scientific team and leverage our advanced technologies to help accelerate the preclinical drug discovery process.

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      • Complex Cell Model Screening Services
        Preclinical services

        Work with our experienced scientific team and leverage our advanced technologies to help accelerate the preclinical drug discovery process.

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      • Base Editing Platform
        • Pin-point base editing platform cell line engineering services
        • Pin-point base editing platform pooled tiled screening services
        Preclinical services

        Work with our experienced scientific team and leverage our advanced technologies to help accelerate the preclinical drug discovery process.

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      • Immune Cell Screening
        Preclinical services

        Work with our experienced scientific team and leverage our advanced technologies to help accelerate the preclinical drug discovery process.

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      • Functional Genomic Screening Services
        Preclinical services

        Work with our experienced scientific team and leverage our advanced technologies to help accelerate the preclinical drug discovery process.

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      • Cell Panel Screening
        Preclinical services

        Work with our experienced scientific team and leverage our advanced technologies to help accelerate the preclinical drug discovery process.

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      • Cell Line Engineering
        • CHOSOURCE CLE Services
        Preclinical services

        Work with our experienced scientific team and leverage our advanced technologies to help accelerate the preclinical drug discovery process.

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      • Viral Vector Engineering and Manufacture
        Preclinical services

        Work with our experienced scientific team and leverage our advanced technologies to help accelerate the preclinical drug discovery process.

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      Preclinical services

      Work with our experienced scientific team and leverage our advanced technologies to help accelerate the preclinical drug discovery process.

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    • Revvity Omics Services
      • Revvity Omics Clinical Services
        • Cytogenomics
        • Global Laboratory Network
        • Metabolic Testing
        • Newborn Screening Services
        • Prenatal Screening Services
        • Proactive Testing
        • Rare Disease Testing
        • Specialized and Customized Assays
        • Sponsored Testing Programs
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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      • Revvity Omics Pharma Services
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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      T-SPOT.TB testing services.

      Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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    • Clinical & Testing Services
      • Revvity Omics Clinical Services
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

        Learn more

      • Revvity Omics Pharma Services
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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      • Cellular and Humoral Immunoassays
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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      • Tuberculosis Testing Services
        • Oxford Diagnostic Laboratories Service Closures
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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      T-SPOT.TB testing services.

      Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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    • Customization Services
      • Assays and Reagents
        • Assay Development
        • Compound profiling
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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      • Microplate Services
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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      • Custom Conjugation & Labeling
        • Custom Labeling
        • Small Molecule
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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      • Radiosynthesis and Labeling
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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      T-SPOT.TB testing services.

      Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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    • Licensing
      • Gene Delivery Licensing
        CHOSOURCE expression platform

        Revvity's expression platform provides an enhanced system for the development and manufacturing of biotherapeutics that can be used in commercial manufacturing applications.

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      • Gene Expression Systems
        • CHOSOURCE Platform
        CHOSOURCE expression platform

        Revvity's expression platform provides an enhanced system for the development and manufacturing of biotherapeutics that can be used in commercial manufacturing applications.

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      • Pin-point™ Base Editing Platform
        CHOSOURCE expression platform

        Revvity's expression platform provides an enhanced system for the development and manufacturing of biotherapeutics that can be used in commercial manufacturing applications.

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      • Virus Screening
        CHOSOURCE expression platform

        Revvity's expression platform provides an enhanced system for the development and manufacturing of biotherapeutics that can be used in commercial manufacturing applications.

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      CHOSOURCE expression platform

      Revvity's expression platform provides an enhanced system for the development and manufacturing of biotherapeutics that can be used in commercial manufacturing applications.

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    • Viral Vector Engineering and Manufacture
      • AAV Services
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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      • Lentivirus Services
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

        Learn more

      T-SPOT.TB testing services.

      Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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    • Instrument Service & Maintenance
      • AV Services
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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      • Equipment Service Plans
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

        Learn more

      • On-demand Equipment Service
        T-SPOT.TB testing services.

        Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

        Learn more

      T-SPOT.TB testing services.

      Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

      Learn more

    • Customer Training
      T-SPOT.TB testing services.

      Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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    • OEM Solutions
      T-SPOT.TB testing services.

      Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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    T-SPOT.TB testing services.

    Revvity's Oxford Diagnostic Laboratories is a large referral laboratory for tuberculosis testing services based on our T-SPOT technology.

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  • Company
    • Purpose
      • Approach
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      • Our Story
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      • Leadership
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    • Careers
      • Careers Home
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      • Why Revvity
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      • Search Jobs
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    • Investor Relations
      • Events
        Investor Day 2024

        Driving meaningful innovation that profoundly impacts science and human lives.

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      • Financials
        Investor Day 2024

        Driving meaningful innovation that profoundly impacts science and human lives.

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      • Stock Info
        Investor Day 2024

        Driving meaningful innovation that profoundly impacts science and human lives.

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      Investor Day 2024

      Driving meaningful innovation that profoundly impacts science and human lives.

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    • ESG
      • Environmental
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      • Social
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      • Governance
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    • News
      • Announcements
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      • Awards
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      • Media Alerts
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      • Press Coverage
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  • Resources
    • Product Support
      • Application Support Knowledge base (ASK)
        Tech documents, at your fingertips.

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      • SpectraViewer
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      • RAD Calculator
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    • Blog
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    • Events
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    • Customer Training
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    • Help Center
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      • Cell Line Terms & Conditions
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    • Knowledge Base
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      • Sample homogenization applications and protocols
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      • Newborn screening disorders
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      • TB testing services
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      • Investigator led studies
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      • Tuberculosis (TB)
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  • Brands
      Revvity Brands
    • BioLegend

    • EUROIMMUN

    • Revvity Omics

    • Revvity Signals

    • Tulip Diagnostics

    • ViaCord

    • Popular Revvity Product Brands
    • AlphaLISA and SureFire Ultra

    • BioQule NGS

    • chemagic

    • EnVision Nexus

    • Fontus

    • HTRF

    • IVIS

    • LEGENDplex

    • Opera Phenix

    • T-SPOT

    • TotalSeq

    • View all Revvity brands
  • Welcome to Revvity: renowned brands and boundless innovation.

    Hearing the word "can't" is our call to action!

    We help scientists, researchers, and clinicians overcome the world's greatest health obstacles.

    View our story

    Featured brand: BioLegend

    Learn about our world-class antibodies for a diverse set of research areas including immunology, neuroscience, cancer, stem cells and cell biology.

    Visit BioLegend.com

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  • HTRF Human MMP1 Detection Kit, 500 Assay Points

HTRF Human MMP1 Detection Kit, 500 Assay Points

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HTRF Human MMP1 Detection Kit, 500 Assay Points
Generic HTRF biomarker image
Generic HTRF biomarker image
Generic HTRF biomarker image

The HTRF Human MMP1 Detection Kit enables the specific and detection and quantification of MMP1 in human cell culture supernatants.

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Feature Specification
Application Protein Quantification
Sample Volume 16 µL
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The HTRF Human MMP1 Detection Kit enables the specific and detection and quantification of MMP1 in human cell culture supernatants.

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Product variants
Unit Size: 500 assay points
Part #:
62MMP1PEG
List price
USD 1,670.33
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Unit Size: 10,000 assay points
Part #:
62MMP1PEH
List price
USD 19,683.38
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For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).
Total list price:
USD 1,670.33
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Generic HTRF biomarker image
HTRF Human MMP1 Detection Kit, 500 Assay Points
Generic HTRF biomarker image
Generic HTRF biomarker image
HTRF Human MMP1 Detection Kit, 500 Assay Points
Generic HTRF biomarker image
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Product information

  • Overview
  • How it works
  • Assay details
  • Analytical performance
  • Assay validation
  • Simplified pathway
  • Specifications

Overview

Matrix metalloproteinase (MMP) 1, also known as interstitial collagenase, is an endopeptidase secreted as an inactive proenzyme that is then activated by proteolytic cleavage. MMP1 plays a significant role in tissue repair and remodeling by degrading structural components of the extracellular matrix (ECM), such as collagens, gelatin, and fibronectin. As such it is closely related to all matrix-dependent disorders, such as fibrotic disease, and some types of cancer that rely on cell adhesion, migration, and invasion.

How it works

Human MMP1 assay principle

The Human Matrix MetalloProteinase 1 (MMP1) assay measures MMP1 in cell supernatant. The assay uses two labeled antibodies: one coupled to a donor fluorophore, the other to an acceptor. In presence of Human MMP1 in a sample, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor, and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the capsid present in the sample and therefore provides a means of assessing any changes caused by experimental variables, in a no-wash assay format.

Principle of the HTRF Human MMP1 assay
Human MMP1 assay protocol

The Human MMP1 assay protocol, using a 384-well small volume white plate,  is described on the right. 16 µL of sample or standard are dispensed directly into the detection plate for detection by HTRF® reagents. The antibodies labeled with HTRF fluorophores are added before an overnight incubation. The assay can be run in up to a 1536-well format by simply resizing each addition volume proportionally.

Protocol of the HTRF Human MMP1 assay

 

Assay details

Human MMP1 assay details
Human MMP1 standard curve
Sample size 16 µL
Final assay volume 20 µL
Time to result Overnight at RT
Detection Limit (LOD) in Diluent 46.5 pg/mL
Dynamic Range 181 – 50 000 pg/mL
Species Human only

 

Analytical performance

Intra-assay

Each of the 3 samples was measured 24 times, and the % CV was calculated for each sample.

Samples were cell culture supernatants from unstimulated MRC-5 cells.

Sample

Mean [Human MMP1] (pg/mL)

CV

1

9 830

3%

2

3 280

4%

3

1 090

5%

 

Mean CV

4%

Inter-assay

Each of the samples was measured in 4 independent experiments by 4 different operators, and the % CV was calculated for each sample. Samples were cell culture supernatants from unstimulated MRC-5 cells.

Sample

Mean [Human MMP1] (pg/mL)

CV

1

6 710

4%

2

3 430

7%

3

1 720

10%

 

Mean CV

7%

Dilutional linearity

Samples were from  primary human lung fibroblast supernatants serially diluted in cell culture medium.

The recovery % obtained from these experiments show the good dilutional linearity of the assay (dilution test acceptance criteria: 85-115%).

Dilution Factor

[MMP1] Expected (pg/mL)

[MMP1] Measured (pg/mL)

Dilution Recovery

Neat

-

49 913

100%

2

24 956

26 783

107%

4

12 478

13 819

111%

8

6 239

6 682

107%

16

3 120

3 031

97%

 

Mean CV

 

104%

Antigen spike and recovery

Four concentrations of recombinant protein (~2815 – 1268 – 546 and 164  pg/mL) were added to 2 dilutions of native sample from primary human lung fibroblast supernatants, and the expected concentrations were compared to those measured in order to compute antigen recoveries (acceptance criteria: 85-115%). The recovery was good after overnight incubation at room temperature. The 100% recovery indicates similar measurements of MMP1 from samples and the kit standard.

Sample

[MMP1] Standard(pg/mL)

[MMP1]Spiked Sample (pg/mL)

Expected (pg/mL)

Measured (pg/mL)

Recovery

1

2 815

9 700

12 516

13 435

107%

1 564

4 379

4 652

106%

2

1 268

9 700

10 969

11 600

106%

1 564

2 832

2 999

106%

3

546

9 700

10 246

11 200

109%

1 564

2 110

2 239

106%

4

164

9 700

9 864

10 993

111%

1 564

1 728

1 940

112%

 

Mean CV

 

 

 

108%

Interferences

The possible interference from human Tissue Inhibitors of Metalloproteinases (TIMP-1, TIMP-2, TIMP-3, and TIMP-4) and α2-macroglobulin was investigated. The human MMP1 was kept at a constant concentration (25 000 pg/mL). The binding proteins were titrated into the assay. No significant interference was observed up to 100 000 pg/mL, which was the maximum concentration tested.

Tested Protein

Interferences (%)100 000 pg/mL

TIMP1

No

TIMP2

No

TIMP3

No

TIMP4

No

α2-macroglobulin

-18%

Cross reactivities

Cross reactivities were assessed using recombinant proteins from the MMP family. Proteins were tested up to 30.000 pg/mL and standard curves were generated for each protein diluted in the kit diluent.

16 µL of sample were transferred into a white detection plate (384 low volume), and 4 µL of a mixture of the HTRF Human MMP1 detection reagents were added. The HTRF signal was recorded after an overnight incubation at room temperature.

Signals were plotted on the assay standard curve to interpolate concentrations. Cross-reactivity was calculated as a mean of 6 tested concentrations. The assay is human specific and mouse MMP1 was not detected by it.

Tested Protein

Cross Reactivity

Human MMP1 (Pro & Active forms)

100%

Mouse MMP1

No

Human MMP2

No

Human MMP3

No

Human MMP8

No

Human MMP9

No

Human MMP13

No

 

Assay validation

MMP1 upregulation induced by different treatments in human pulmonary fibroblast cell line MRC-5

The human lung fibroblast cell line MRC-5 was plated in a 96-well plate (12,500 cells/well). The day after, cells were treated overnight with 50 µL of increasing concentrations of PMA diluted in FBS-free medium supplemented with 1% BSA. Cell supernatants were collected for the quantitative measurement of secreted MMP1 levels.

16 µL of supernatants (diluted ¼) were then transferred into a white detection plate (384 low volume) and 4 µL of the HTRF Human MMP1 detection reagents were added. The HTRF signal was recorded after an overnight incubation at room temperature.

As expected, PMA treatment led to a dose-dependent increase in the secretion of human MMP1.

The human lung fibroblast cell line MRC-5 was plated in a 96-well plate (12,500 cells/well). The day after, cells were treated overnight with 50 µL of increasing concentrations of human TNF alpha or TGF-β1 diluted in FBS-free medium supplemented with 1% BSA. Cell supernatants were collected for the quantitative measurement of secreted MMP1 levels.

16 µL of supernatants (diluted ¼) were then transferred into a white detection plate (384 low volume) and 4 µL of the HTRF Human MMP1 detection reagents were added. The HTRF signal was recorded after an overnight incubation at room temperature.

As expected, cell treatment for 24h induced an upregulation of the secreted level of human MMP1, particularly after TNF alpha treatment.

Validation of HTRF Human MMP1 kit on MRC-5 cell line
Validation of HTRF Human MMP1 kit on MRC-5 cell line
MMP1 upregulation induced by PMA treatment in cell lysates and cell supernatant of human glioblastoma cancer cell line U87-MG

The human glioblastoma cancer cell line U87-MG was plated in a 96-well plate (100,000 cells/well). The day after, cells were treated overnight with 50 µL of PMA (50 ng/mL) diluted in FBS-free medium supplemented with 1% BSA.

Cell supernatants were collected, and cells were lysed with 50 µL of supplemented lysis buffer #3 (4X) for 30 min at room temperature under gentle shaking for the quantitative measurement of MMP1 levels.

16 µL of supernatants or lysates were then transferred into a white detection plate (384 low volume) and 4 µL of the HTRF Human MMP1 detection reagents were added. The HTRF signal was recorded after an overnight incubation at room temperature.

As expected, PMA treatment led to an increase in the intracellular and secreted levels of human MMP1 in this cancer cell line.

Validation of HTRF Human MMP1 kit on U87-MG cell line in cell lysates and supernatant
Versatility of Human MMP1 assay in human cell line supernatants

Different human cell lines were seeded in T175 flasks in 25 mL of complete culture medium at 37°C, 5% CO2 up to 80% confluence. Then the supernatant was collected. 16 µL of each cell line supernatant were transferred into a 384 well low volume white microplate and 4 µL of HTRF Human MMP1 detection antibody were added. The HTRF signal was recorded after an overnight incubation. The results reveal a differential expression pattern with a high concentration level in the human pulmonary fibroblast cell line and a lower level in the human glioblastoma cancer cell line (U87-MG) or Human foreskin BJ fibroblasts.

Assessment of Human MMP1 level in supernatant of various cell lines

 

Simplified pathway

MMP1 signaling pathway

Matrix metalloproteinases (MMPs) are a family, 23 members of which have been identified, produced by diverse cell types, including immune cells, epithelial cells, and fibroblasts. MMP functions include degradation of components of the extracellular matrix (ECM) in normal physiological processes, such tissue remodeling, and in disease processes, such as cancer. Specifically, MMP-1 breaks down the interstitial collagens, types I, II, and III.

MMP1, or collagenase-1, plays critical roles in many physiologic processes by degrading these collagen types I, II, and III.

The regulation of MMP1 is complex and multifactorial. All the pathways involved seem to converge toward JNK and its resulting activation of transcription factors c-Jun and c-Fos. Various growth factors and cytoskeleton-related mechanical tension may be involved.

Cytokines are also involved in MMP1 regulation. Pro-inflammatory IL-1β in particular promotes them via the NF-kB pathway or via p38 MAP kinase, particularly in cancer.

MMP1 are secreted or expressed at the membrane as precursors and their activity is regulated by the TIMPs (tissue inhibitors of metalloproteinases) and by themselves, as they activate each other in a complex regulatory network.

MMP1 signaling pathway

 

Specifications

Application
Protein Quantification
Brand
HTRF
Detection Modality
HTRF
Product Group
Kit
Sample Volume
16 µL
Shipping Conditions
Shipped in Dry Ice
Target Class
Biomarkers
Target Species
Human
Technology
TR-FRET
Unit Size
500 assay points

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    SDS IconSafety Data Sheet (US) : HTRF Human MMP1 Detection Kit / 62MMP1PEG
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    SDS IconSafety Data Sheet (FRA) : HTRF Human MMP1 Detection Kit / 62MMP1PEG
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    SDS IconSafety Data Sheet (DEU) : HTRF Human MMP1 Detection Kit / 62MMP1PEG
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    SDS IconCertificate of Analysis : HTRF Human MMP1 Detection Kit / 62MMP1PEG
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    SDS IconManual : HTRF Human MMP1 Detection Kit / 62MMP1PEG-62MMP1PEH

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