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WORKFLOW KIT · TARGET ENGAGEMENT
Measure intracellular target engagement in live cells, no digitonin, no lysis.
Kits include: Custom cargoes · Optimized protocols · Portal reagents · Delivery cartridges · Suggested readouts
Run the NanoBRET target engagement assay you already use, on cells that are alive and intact. One pass through a MicroBooster™ cartridge puts an impermeable tracer and the compound into the cytosol together, so you read binding where the target actually sits. No digitonin, no lysis, and no permeable analogue to design first.
The Current Workflow is Broken
Target engagement is an intracellular question, and the tracers and probes that answer it are often the ones unable to get there. Every standard approach costs you something. Digitonin permeabilization and lysate assays let the reagent in by giving up the live cell, which costs the target's cellular context and returns a single endpoint instead of kinetics. Engineering your way around it costs time and money.
01
Lyse or permeabilize cells
02
Run target engagement assay
03
Lose live-cell context
Dead End
Portal has the Solution
01
Prepare
NanoLuc-tagged POI cells
02
Mix
Cells + tracer and compound
03
Boost
Gateway™ mechanoporation
04
Incubate
Let the signal develop
05
Measure
Live-cell NanoBRET signal
Target engagement measured in live cells
~20×
Live-cell BRET from a boosted impermeable Src tracer, against the same tracer unboosted
2.3×
Ubiquitination response for partially permeable degraders once delivered, in intact cells
80+%
Delivery held across ten consecutive 96-well plates of HeLa cells
The cargo and protocols below make this kit specific. The core consumables, settings, and support ship with every Portal kit.
Available Cargoes
Reagents & Consumables
Protocol & Settings
Support
Available Cargoes
Reagents & Consumables
Protocol & Settings
Support
We boosted a membrane-impermeable Src kinase tracer into HeLa cells expressing NanoLuc-Src, then challenged it with ponatinib or bosutinib. Boosted cells gave about 20× the BRET signal of unboosted cells. Ponatinib displaced the tracer dose-dependently, cutting the signal about 35% at 1.1 µM, about 55% at 3.3 µM, and about 80% at 10 µM; bosutinib reached about 85% at 10 µM. Unboosted cells stayed at baseline across the whole curve.
Cellular ubiquitination of the target protein in a degrader mechanism-of-action panel run by a pharma partner, showing normalized POI:ubiquitin BRET response. Both partially permeable compounds increased ubiquitination about 2.3-fold after delivery, while DMSO showed no significant difference and the permeable control was unchanged. Ternary complex formation and proteasome recruitment increased to the same extent in the same panel.
Anti-phospho-BTK detection antibodies boosted into human PBMCs and read on a luminescent immunoassay. After pervanadate stimulation, the delivered-antibody arm reached about 80% of the lysate signal, with no-cargo controls near zero: pBTK signal mirrors lysate without permeabilization. Assay chemistry: Promega Lumit.
Percent delivery for every well across ten consecutive 96-well plates of HeLa cells, boosted in line with the Powered-by-Portal Galaxy-i consumable on a Certus Flex dispenser. Delivery started near 100% and settled in the high 80s, holding between about 80% and 100% for the whole run. How much tracer gets in sets the BRET ratio, so this consistency keeps dose-response curves comparable from plate to plate. The approach scales directly to high-throughput screening on the Galaxy-i.
mRNA, siRNA, saRNA
Proteins & Peptides
CRISPR RNPs
Small Molecules
Polymers
Nanoparticles
Antibodies
Virus
PBMCs
T cells
B cells
NK cells
iPSCs
Monocytes
RBCs
HSCsmRNA, siRNA, saRNAProteins & PeptidesCRISPR RNPsSmall MoleculesPolymersNanoparticlesAntibodiesVirusPBMCsT cellsB cellsNK cellsiPSCsMonocytesRBCsHSCsValidated live-cell workflows for target engagement, compound profiling, and beyond.
Every kit includes: Cargoes · Optimized protocols · MicroBooster™ cartridges · Suggested readouts · Reagents
