Issue link: https://resources.nanotempertech.com/i/1531548
1 4 A P P L I C A T I O N N O T E Highlights No Need for Immobilization: Monolith X, when paired with the NativeMP™ Platform, ensures mem- brane proteins are stabilized within their natural lipid environment, maintaining their authentic structure and dynamics without the need for detergents. This approach eliminates the challenges of immobilization, such as steric hindrance or conformational changes, ensuring binding pockets remain accessible and unaltered. Native-Like Environment Compatibility: Monolith X uses advanced Spectral Shi and TRIC tech- nologies for precise binding measurements directly in solution. These techniques detect binding events through fluorescence changes rather than mass shi s, making them highly effective for large protein complexes. This approach ensures compatibility with native-like lipid or copolymer environ- ments, enabling accurate analysis of membrane protein interactions under near-native conditions. Small Sample Requirement: The Monolith X system requires minimal sample amounts, a signi- ficant advantage for membrane proteins that are o en challenging to prepare and yield in limited quantities. This efficient use of resources makes it an ideal solution for studying rare or delicate targets, as demonstrated in complex interactions like NTCP and LHBsAg, where high analyte yields could otherwise pose purification challenges. No Mass Disparity Sensitivity: Monolith X excels in detecting interactions across a wide range of molecular weights by leveraging fluorescence-based sensitivity. This capability allows it to resolve binding events between large protein complexes and small molecule analytes, such as the P2X4 receptor (150 kDa in its trimeric form with additional lipid mass) and 5-BDBD (355 Da). Even with extreme mass differences, Monolith X delivers precise and reliable results, overcoming limitations typically faced in mass-dependent detection systems. Fast Turnover Times: Monolith X delivers rapid results, a measurement of a dilution series takes no longer than 6 minutes combining Spectral Shi and TRIC. This efficiency is ideal for studying tight binders or hydrophobic small molecules prone to aggregation in aqueous solutions. The shorter processing time improves data quality and increases the success rate of binding studies.