Description
Key facts
Detection method:Fluorescent,
Sample types:Tissue Homogenate, Cell Lysate,
Assay type:Enzyme activity (quantitative),
Assay Platform:Microplate
Product details:
The Glucosylceramidase Activity Assay Kit (Fluorometric) is a sensitive assay to measure the the activity level of the glucosylceramidase enzyme in a sample by quantifying the fluorescence intensity of a cleaved synthetic substrate in tissue homogenates & cell lysates.
How the assay works
Glucosylceramidase Activity Assay Kit (Fluorometric) uses a synthetic substrate containing a fluorescent molecule, where the glucosylceramidase enzyme cleaves the substrate, releasing the fluorescent molecule which can then be quantified by measuring its fluorescence intensity (Ex/Em=360/445 nm), thus allowing for the determination of the enzyme's activity level in a sample; essentially, the more fluorescent signal detected, the higher the glucosylceramidase activity present.
Assay protocol summary
Glucosylceramidase Activity Assay Kit (Fluorometric) protocol summary
- Prepare all reagents, samples and positive controls.
- Add all samples to the appropriate wells.
- Prepare and add Glucosylceramidase Substrate to the appropriate wells.
- Measure fluorescence y (Ex/Em = 360/445 nm) at 37°C in end point mode
- Determine Glucosylceramidase activity using equation.
This product is manufactured by BioVision, an Abcam company and was previously called K2003 Glucosylceramidase Activity Assay Kit (Fluorometric). K2003-100 is the same size as the 100 test size of ab273339.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
GBA also known as glucosylceramidase is a lysosomal enzyme with a molecular mass of approximately 59 kDa. This enzyme breaks down glucosylceramide into glucose and ceramide. GBA is expressed predominantly in tissues with high metabolic activities such as the brain liver and spleen. Its function relies on its catalytic activity where substrates bind to its active site enabling the hydrolysis process necessary for maintaining cellular metabolism.
Biological function summary
GBA plays an important role in sphingolipid metabolism. It participates in the degradation of glycolipids within the lysosome contributing to lipid recycling. It acts independently rather than as a part of a major enzymatic complex. Through its role in degrading glucosylceramide GBA influences cellular homeostasis and bioenergetics ensuring balance in neural and systemic lipid levels.
Pathways
GBA’s enzymatic functions are integral to the glycosphingolipid metabolic pathway. It is involved in the downstream steps of the lysosomal degradation of glycosphingolipids. The pathway operates alongside other important proteins such as beta-glucosidase and CERT-related transfer proteins all of which contribute to membrane lipid organization and signal transduction processes.
GBA mutations are linked with Gaucher disease and Parkinson’s disease. In Gaucher disease deficient GBA activity leads to substrate accumulation resulting in hepatosplenomegaly and other systemic symptoms. Reduced GBA activity is also associated with increased alpha-synuclein aggregation in Parkinson’s disease implicating it in the pathogenesis of neurodegenerative disorders. The enzyme’s function in these diseases highlights its role in maintaining cellular equilibrium and signaling pathways.
