Buffers with ß-Glucuronidase Substrate
The b-Glucoronidase (GUS) reporter assay system is one of the most used reporter gene for plant gene expression reseach, to study gene regulation events and for evaluation transient and stable transformations in plants. b-Glucoronidase is found in plant and mammalien cells. The human b-Glucuronidase is found in lysosomes and is homologe to the Gene from E.coli (uidA) which is used as reporter. This effectiveness is caused by the enzyme stability, high sensitivity and suitabbility for the assays to detection by fluorometric, spectrophotometric, histochemical and chemiluminescent techniques.
PJK´s series of stabilized chemiluminescent 1,2-dioxetanes substrates can detect ß-Galactosidase enzyme at low levels:
- Reporter Gene Assays
Store the reagent bottles at 2-6 °C or at -20 °C.
Equilibrate the reagent at room temperature till clear solution observed if stored at -20 °C.
Do not contaminate the ?-Glucuronidase substrate with ?-Glucuronidase enzyme or other proteins.
?-Glucuronidase substrate has a wide range to detect ?-Glucuronidase enzyme in solution as well as on membrane. The lower enzyme concentration may be atto grams and the highest concentration of ?-Glucuronidase enzyme may be nano gram depending on the source of the ?-Glucuronidase enzyme.
For good results, wash your tube or micro titer plate or membrane with 0.1M phosphate buffer, pH 7.2 to 7.4, and then use the substrate.
Best results for chemiluminescence detection of ?-Glucuronidase enzyme or reporter assays can be obtained from 30 minutes to 45 minutes incubation of ?-Glucuronidase substrate with ?-Glucuronidase enzyme and read the plate or tube within 2 to 10 minutes after triggering the reaction mixture by accelerator or enhancer.