PLCglow™ is the only soluble, fluorescent reporter of phospholipase C (PLC) isozymes that will lead to smarter, safer and more effective drug screening and in the biochemical research of a target.
Unique properties include:
- Soluble, fluorescent reporter of phospholipase C (PLC) isozymes.
- Specifically designed for mammalian PLCs:
- PLCglow™ was specifically designed to reflect the endogenous substrate, phosphatidylinositol 4,5-bisphosphate( PIP2). The two substrates produce similar values of Km and Vmax with various mammalian PLCs.
- Will also work with the more promiscuous bacterial PLCs.
- Robust output.
- Greater than six-fold change in fluorescent when hydrolyzed by PLCs.
- Works with all enzymatically active PLC isozymes.
- Will not report activities of other phospholipases (PLD, PLA2).
- Works with purified PLCs and cellular lysates.
- No comparable reagent on the market.
- Replaces tedious metabolic labeling with radioactive inositol to measure PLCs.
- Stable up to a year at 80°C or 3 weeks upon re-suspension in water.
PLCglow™ is a small molecule that directly reports the enzymatic activity of phospholipase C (PLC) isozymes1,2. PLCglow™ is hydrolyzed by PLCs to generate inositol trisphosphate (IP3), quinomethide, and 6-aminoquinoline. Relative to PLCglow™ , the aminoquinoline is highly fluorescent (λex/em = 344/530 nm) (Fig. 1).
1. Huang et al., A fluorogenic, small molecule reporter for mammalian phospholipase C isozymes. ACS Chemical Biology 6(3), 223-8 (2011).
2. Huang et al., Small molecule inhibitors of phospholipase C from a novel high-throughput screen. JBC 288(8), 5840-8 (2013).
Molecular weight: 1127 g/mol as triethylamine salt (824 g/mol for compound alone).
Amount: 50 μg.
Storage and stability:
Storage temperature: -80°C.
Stability: Stable for up to 12 months at -80°C and up to 14 days at 4°C in water. PLCglow™ is not stable for long periods in several common buffers and stock solutions should be stored only in water. Repeated freezing and thawing is not recommended.
Notes: Results will vary depending on the sensitivity of the fluorometer and the specific activity of the PLC isozyme. In order to obtain optimal results, we recommend testing a dilution series of PLCglow™ and PLC isozyme. PLCglow™ is designed to work with purified proteins and cellular lysates. In contrast, PLCglow™ will not enter intact cells and is therefore inappropriate for measuring intracellular PLCs.