PLCglow™, also known as WH-15, is a unique soluble, fluorogenic reporter of phospholipase C (PLC) isozymes for smarter, safer and more effective biochemical studies. It makes tedious metabolic labeling with radioactive inositol redundant. PLCglow™ directly reports the enzymatic activity of PLC isozymes.1,2 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).
- reflects the endogenous substrate, phosphatidylinositol 4,5-bisphosphate (PIP2)
- is specifically designed for mammalian PLCs, but also works with the more promiscuous bacterial PLCs
- shows greater than six-fold change in fluorescence when hydrolyzed by PLCs
- works with all enzymatically active PLC isozymes, whether purified or cellular lysate
- does not report activities of other phospholipases (PLD, PLA2).
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.