A nanoluciferase (NLuc) sequence is fused to a tetraspanin scaffold such as CD9 or CD63. This directs the reporter to the inner face of EV membranes. Cells stably expressing the construct secrete NLuc-positive EVs. Conditioned media or recipient-cell lysate luminescence, directly proportional to EV quantity, is measured on a standard plate luminometer.
Extracellular Vesicles / Exosomes
Exosome Secretion Inhibitors and Release Modulation Tools for EV Biogenesis Studies
NLuc-based EV reporter tools and secretion modulators for quantifying exosome release and uptake
Reagents that selectively modulate EV secretion let you distinguish exosome-mediated effects from direct cell contact or soluble factor signalling. This helps when studying biogenesis or intercellular cargo transfer. Bioluminescent reporter systems fuse a nanoluciferase tag to EV surface markers such as CD9, CD63 or CD81. This enables quantification of defined EV subpopulations, giving you a real-time readout of secretion rates and recipient-cell uptake on any standard plate luminometer. Luminescence in conditioned media correlates directly with EV secretion, while signal in recipient cells measures transfer.
Three reagent formats support your workflows. Lentiviral expression vectors build stable EV reporter cell lines. Pre-built stable HEK293 reporter lines give you an immediate starting point. Purified EV reporter preparations add directly to recipient cells for uptake studies. EV-Luminite reagents are available with NLuc fused to CD9, CD63 or CD81. This gives flexible labelling and quantification of distinct EV subpopulations for your biological question. Secretion modulation reagents allow pharmacological inhibition or stimulation of EV biogenesis for mechanistic studies.
Choose your exosome release or transfer reagent by experimental question.
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Measuring secretion rate. If you need quantitative EV secretion measurement without nanoparticle tracking analysis, the NLuc-CD9/CD63 or CD81 lentiviral vector suits this. It creates your stable reporter line. Conditioned-media luminescence is then measured with any plate luminometer.
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Measuring recipient-cell uptake. To track transferred EVs in target cells, purified NLuc-EV reporter preparations are added directly to recipient cells. This gives luminescence proportional to EV uptake.
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Speed of setup. For an immediate experiment, a pre-built NLuc-CD9/CD63 or CD81 reporter HEK293 stable cell line is ready to use. For your own-cell reporter, transduction with the lentiviral vector and selection under antibiotic pressure takes two to three weeks to establish.
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Secretion modulation studies. Combining the reporter system with reagents that affect EV secretion, such as GW4869 or bafilomycin A1, quantifies their impact on your EV output. The luminescence readout is more sensitive and higher-throughput than NTA for your dose-response studies.
Applications
Secretion kinetics
NLuc-tagged, tetraspanin-positive vesicles quantify EV secretion from your producer cells. Cross-checking against CD63, CD81 and CD9 quantification ELISAs confirms the result.
Recipient-cell uptake
The same luminescent reporter system measures dose-dependent EV uptake in your specific recipient cell types, distinguishing cargo transfer from surface binding alone.
Intercellular communication mapping
The reporter system maps which producer and recipient cells exchange EV cargo in your co-culture systems. Fluorescent EV labelling kits add visual confirmation.
Secretion pathway inhibitor studies
GW4869 or bafilomycin A1 helps dissect your endosome pathway. The luminescence readout is a sensitive, higher-throughput alternative to NTA.
Uptake receptor identification
Combining the reporter assay with receptor-blocking antibodies or genetic knockdown identifies surface receptors that mediate EV internalisation in your system.
High-throughput screening
Inhibitor or compound screens run on a standard plate luminometer, avoiding outsourced NanoSight analysis. This suits your lab without dedicated tracking kit.
Product Catalog
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Exosome Release & Transfer
NLuc Reporter Tools for Exosome Secretion & Uptake
Frequently asked questions
GW4869 inhibits neutral sphingomyelinase 2, reducing ceramide-dependent multivesicular body biogenesis. Bafilomycin A1 blocks vacuolar-type H-ATPase and disrupts late-endosome acidification. Dimethyl amiloride inhibits Na-H exchangers, affecting membrane trafficking. Each inhibitor targets a different step of the secretory pathway. Using multiple agents helps confirm pathway specificity.
Acid wash or trypsin treatment after incubation removes surface-bound EVs, leaving only the internalised signal. Comparing luminescence or fluorescence before and after acid wash quantifies the internalised fraction. Co-localisation of labelled EVs with endosomal markers such as Rab5 or LAMP1 in confocal imaging further confirms active internalisation versus passive surface attachment.
Include a no-EV vehicle control to establish baseline recipient-cell signal. Also include heat-inactivated EVs (65 °C for 30 min) to distinguish cargo-dependent effects from membrane contact alone. Finally, measure fluorescence or luminescence signal in producer-cell-free conditioned media, to confirm that signal originates from EVs and not secreted soluble proteins.
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