This Human NF-kB assay kit is an all-inclusive firefly luciferase reporter assay system that includes in addition to NF-kB Reporter Cells, optimized media for use during cell culture and in diluting user’s test samples, a positive control activator, Luciferase Detection Reagent, and a cell culture-ready assay plate.

NF-kB Reporter Cells are prepared using INDIGO’s proprietary CryoMite™ process. This cryo-preservation method yields high cell viability post-thaw, and provides the convenience of immediately dispensing healthy, division-competent reporter cells into assay plates. There is no need for intermediate spin-and-wash steps, viability determinations, or cell titer adjustments.

INDIGO’s NF-kB assay kits feature a luciferase detection reagent specially formulated to provide stable light emission between 5 and 90+ minutes after initiating the luciferase reaction. Incorporating a 5-minute reaction-rest period ensures that light emission profiles attain maximal stability, thereby allowing assay plates to be processed in batch. By doing so, the signal output from all sample wells, from one plate to the next, may be directly compared within an experimental set.

NF-kB assay kits are offered in different assay formats to accommodate researchers’ needs: 3x 32, 1x 96, and 1x 384 assay formats for screening small numbers of test compounds, as well as custom bulk reagents for HTS applications.

Assay Kit & Platforms

Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.

Assay Services

The primary application of INDIGO’s cell-based nuclear receptor assays are to quantitatively assess the bioactivity of a test compound as an agonist (activator) or antagonist (inhibition of an agonist response) of a given receptor. Service assays include a positive control reference compound and ‘vehicle’ control for every experiment. A formal study report and all data files are provided to the client upon completion of the study. To receive a quote for your proposed study, complete & submit the online Request a Quote form or contact an INDIGO Customer Service Representative to discuss your desired study parameters.

Background

NF-kB is a signal transduction dependent transcription factor.

INDIGO's Human NF-kB Reporter Assay System utilizes proprietary human cells that express NF-kB (nuclear factor kappa-light-chain enhancer of activated B Cells), and contain the luciferase reporter gene functionally linked to upstream NF-kB genetic response elements. Thus, quantifying changes in luciferase expression provides a sensitive surrogate measure of changes in the level of NF-kB activation.

This NF-kB reporter cell line is validated to provide a robust dose-dependent activation response when treated with TNFa, or the Protein Kinase C activator Phorbol 12-myristate 13-acetate (PMA). As such, the principle application of this assay system is in the screening of test samples to quantify any functional activities that they may exert to modulate, either induce or suppress, NF-kB activities.