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WORKFLOW KIT · LIVE-CELL DETECTION
Measure intracellular proteins in live cells. No lysis, and no tagging of the target.
Kits include: Custom cargoes · Optimized protocols · Portal reagents · Delivery cartridges · Suggested readouts
Read an endogenous intracellular protein, a phosphorylated kinase like p-BTK, while the cells are still alive, with no lysis and no engineered tag. Portal boosts the detection antibodies straight into the cytosol, so signal-to-noise matches a lysate immunoassay while the stimulated versus untreated window is preserved in live cells. The same cells are still there for whatever you measure next.
The Current Workflow is Broken
Reading an intracellular protein normally means lysing the cells, and that ends the experiment. You lose the time course, the single-cell resolution, and every phenotype you would have measured next. The live-cell alternative is to knock a tag into the protein first, which is weeks of cell-line work and is out of reach for most primary cells.
01
Lyse cells
02
Run immunoassay
03
Lose live-cell context
Dead End
Portal has the Solution
01
Prepare
Cells of choice
02
Mix
Cells + intracellular antibodies
03
Boost
Gateway™ mechanoporation
04
Incubate
Allow signal to develop
05
Measure
Luminescent readout in live cells
Measure protein detection 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 anti-phospho-BTK detection antibodies into human PBMCs in one pass and drove BTK phosphorylation with pervanadate. The stimulated live cells reached about 80% of the lysate signal and had a signal-to-noise ratio similar to lysate, so a phosphorylation response can be read out in living cells rather than in a lysate. Assay chemistry: Promega Lumit.
Antibodies are one impermeable detection reagent among several. We boosted LgBiT protein into cells carrying HiBiT knocked into the endogenous KRAS(G12C) locus, where it complements the tag and lights up the protein in place. Adding the G12C-specific degrader LC-2 for 24 hours cut that signal to about 40% of the no-degrader baseline at 1 µM, about 25% at 5 µM, and under 10% at 10 µM. Untreated, no-boost, and no-cargo wells sat at background. The protein level was followed in the same live cells across those 24 hours, with no lysis timepoint in the series. Assay chemistry: Promega HiBiT/LgBiT.
A p-BTK readout is only meaningful if the antibodies reach the cells that carry the signal. BTK is the B cell receptor kinase, so a p-BTK readout in a mixed PBMC prep is a readout of the B cell compartment. We boosted mixed PBMCs with a fluorescent dextran and gated on CD19⁺ cells afterwards. About 86% of live CD19⁺ cells came back cargo-positive against roughly 0% for unboosted control, so cargo reaches the very cells the assay depends on. Viability of the CD19⁺ fraction was about 62% after the boost against about 88% untreated.
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
