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Nuclear Receptor & In Vitro Toxicology Solutions™

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  • ABOUT
    • About INDIGO
    • Why INDIGO
    • Key Personnel
      • Board of Directors & Advisors
      • Management
    • Employment
    • INDIGO Press Releases
    • INDIGO in the News
  • ASSAYS
    • By Receptor
    • By Disease State
      • Overview
      • Anemia & Kidney Disease
      • Autoimmune Disease & Inflammation
      • Cancer
      • Cardiovascular Disease
      • Diabetes
      • Endocrinology
      • NAFLD/NASH
      • Obesity
    • Toxicology Solutions
      • In Vitro Toxicology Platform
      • Gene Expression
      • MDR1 / Human P-Glycoprotein
      • Discovery Toxicology
      • Environmental Monitoring
    • INDIGlo Luciferase Detection Reagent
    • Live Cell Multiplex
    • Ortholog Assays
    • Custom Assay Development
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Live Cell Multiplex Assay Kits & Services

Product Family Product Number Product Description Technical Manual
LCMA LCM01 LCMA 1×96-wells Reagent Pack Technical Manual
LCM05 LCMA 5×96-wells Reagent Pack Technical Manual
LCM10 LCMA 10×96-wells Reagent Pack Technical Manual

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  • Format & Performance

The Live Cell Multiplex (LCM) Assay provides an efficient fluorescence-based method of quantifying the relative number of live cells resident in treated wells of an assay plate. While the LCM Assay may be performed as a stand-alone assay, it has been specifically optimized to be run in multiplex with any of INDIGO’s 96-well, 2x48-well, or 3x32-well Nuclear Receptor Reporter Assay System products.

The LCM assay allows users to validate their primary Nuclear Receptor Assay data by determining if their test compound treatments exert mitogenic, cytostatic or cytotoxic activities on the reporter cells. The occurrence of such adverse non-specific effects will always undermine the accurate assessment of a test compound’s potency and/or efficacy as a modulator of nuclear receptor function.

When screening test compounds for antagonist activities it is particularly important to quantify changes in the relative number of live reporter cells at the assay endpoint. Test compounds that exert cytostatic, cytotoxic, or cytolytic activities invariably generate “false-positive” results in an antagonist screen. In such cases, the observed drop in luciferase activity will be incorrectly attributed to inhibition of the nuclear receptor by the test compound. In reality, however, the treatment compound has pushed the reporter cells into division arrest, apoptosis, necrosis, or lysis.

The LCM assay allows user’s to validate their primary Nuclear Receptor Assay data by determining if their test compound treatments exert mitogenic, cytostatic or cytotoxic activities on the reporter cells. The occurrence of such adverse non-specific effects will always undermine the accurate assessment of a test compound’s potency and/or efficacy as a modulator of nuclear receptor function.

When screening test compounds for antagonist activities it is particularly important to quantify changes in the relative number of live reporter cells at the assay endpoint. Test compounds that exert cytostatic, cytotoxic, or cytolytic activities invariably generate “false-positive” results in an antagonist screen. In such cases, the observed drop in luciferase activity will be incorrectly attributed to inhibition of the nuclear receptor by the test compound. In reality, however, the treatment compound has pushed the reporter cells into division arrest, apoptosis, necrosis, or lysis.

Format

Live Cell Multiplex Format The fluorescence-based LCM Assay and the luminescence-based INDIGO NR Assay are performed sequentially using the same assay wells. The text in blue corresponds to the LCM Assay portion of the multiplex protocol. Text in black corresponds to the standard protocol used for each of INDIGO’s 96-well format Nuclear Receptor Reporter Assays.

Performance

Live Cell Multiplex performance RFU = % Live Cells. The LCM Assay provides a direct correlation between % RFU and % Live Cells in an assay well.

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