WH-15 

WH-15 is a fluorogenic PLC reporter with K_m value of 49; 30, 86.1 µM for PLC-γ1, PLC-δ1, PLC-β2, respectively. WH-15 can be cleaved in a cascade reaction to generate fluorescent 6-aminoquinoline. WH-15 can be used to respond to imaging PLC activity in live cells (Ex/Em = ~344/530 nm)^[1].

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)^[1].
1. Solution preparation
1.1 Preparation of stock solution
Solvent: DMSO
Concentration: 25 mM (optimized according to the experiment).
1.2 Preparation of working solution
Dilute to 50 μM with assay buffer.
Note: The working solution should be prepared and used immediately. Keep it away from light.

2. Cell staining
2.1 HEK-293T cells are transfected with 600 ng of the indicated DNA and 100 ng of empty vector.
2.2 Add 70 μL of 50 μM WH-15 (83 μg/mL fatty-acid free BSA, 50 mM HEPES (pH 7.2), 70 mM KCl, 3 mM CaCl2, 3 mM EGTA and 2 mM DTT) to 50 μL normalized cell lysate at 37°C.
2.3 Data is collected every 3 min using a Multilabel Counter (Ex/Em = 340/535 nm).

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

824.60

C₃₁H₄₃N₂O₁₈P₃

1264748-47-1

Solid

Light yellow to yellow

O[C@@H]1[C@H](OP(O)(OC2=CC=C(COC(NC3=CC=C4N=CC=CC4=C3)=O)C=C2OCCCCCCCC)=O)[C@@H](O)[C@H](OP(O)(O)=O)[C@@H](OP(O)(O)=O)[C@@H]1O

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light, stored under nitrogen

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Huang W, et al. A fluorogenic, small molecule reporter for mammalian phospholipase C isozymes. ACS Chem Biol. 2011 Mar 18;6(3):223-8.  [Content Brief]