Quantification of Live Cell Intracellular Protein Levels with Promega LgBiT and Boosting

Who this is for:

Early-stage drug discovery teams

Opportunities

Detection of intracellular protein levels in live cells: LgBiT delivery allows for live cell detection while current methods require cell lysis, losing cell-specific and time resolution in your assays
No need for LgBiT cell lines: Building constructs and cell selection lengthens timelines
Work with sensitive cell lines: Delivery via mechanoporation can work in primary cells, allowing for more biologically relevant assays

Results Obtained

Intracellular protein detection in live cells with Promega’s HiBiT/LgBiT reagents

How We Did It

Portal uses mechanoporation to deliver virtually any cargo to diverse cell types while maintaining cell health. This enables intracellular delivery of impermeable molecules without the complications of electroporation or other delivery methods. Learn more about mechanoporation here!

Fig 1: Relative Luminescence, 24h PROTAC Treatment. MiaPaCa-2 cells endogenously tagged with HiBiT at the KRAS(G12C) locus were treated with the PROTAC LC-2 for 24 hours (0-10 μM) and mechanoporated with LgBiT protein (Promega) Nano-Glo® Endurazine™ was added and luminescence was detected on a GloMAX plate reader to detect degradation of KRAS(G12C).

Live Cell Intracellular Protein Levels Measurement

Using Portal’s mechanoporation approach we successfully delivered LgBit protein to quantify degradation of KRAS(G12C) tagged with HiBiT by the G12C-specific PROTAC degrader, LC-2 (Fig 1). This approach could also be used in other intracellular labeling assays to determine protein-protein interactions (PPIs), protein stabilization, and other live cell kinetics.