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Citações e referências (16)
Invitrogen™
PFB-FDGlu (5-(Pentafluorobenzoylamino)Fluorescein Di-β-D-Glucopyranoside)
PFB-FDGlu is a cell-permeable substrate used for live cell imaging of GCase activity. It fluoresces green upon cleavage by lysosomal GCase, facilitating specific, single-cell analysis via high-content imaging, flow cytometry, and microscopy.
| Número do catálogo | Quantity |
|---|---|
| P11947 conhecido também como P-11947 | 5 mg |
Número do catálogo P11947
conhecido também como P-11947
Preço (BRL)
-
Quantity:
5 mg
PFB-FDGlu (5-(Pentafluorobenzoylamino) Fluorescein Di-beta-D-Glucopyranoside) is a glycosidase activity probe for live cell imaging of lysosomal glucocerebrosidase (GCase) activity. This enzyme is vital for glycolipid metabolism, and its deficiency is linked to diseases like Gaucher’s and Parkinson's. PFB-FDGlu fluoresces green upon cleavage by GCase, allowing specific, single-cell analysis via flow cytometry, fluorescent microscopy, and high-content imaging.
PFB-FDGlu is cell-permeable and when loaded into cells, localizing to endosomal and lysosomal compartments while maintaining cell viability. Upon enzymatic cleavage by active GCase enzymes, it yields a green-fluorescent dye (PFB-F) with excitation/emission maxima of approximately 492/516 nm. This fluorescence can be detected using fluorescence microscopy, high-content imaging, and flow cytometry, allowing for specific, single-cell analysis of GCase activity.
Advantages of PFB-FDGlu for detecting GCase activity in lysosomes:
- High specificity—utilizes a lysosomal GCase substrate for measurement of GCase activity, ensuring specific lysosomal staining (note that with long incubations the fluorescent PFB-F will migrate to the cytoplasm)
- Live cell imaging—allows for the assessment of GCase activity in intact, living cells with minimal impact on cell viability, even with extended incubation
- Single-cell analysis—enables specific, single-cell analysis with flow cytometry, high-content imaging, and microscopy to distinguish between different cell types
- Bright fluorescence—yields a bright green fluorescent dye upon cleavage by GCase, enhancing detection sensitivity
- Versatile protocol—can be combined with other phenotypic indicators, such as viability stains and antibodies, to identify distinct cell populations or as a selection criterion for cell sorting
GCase, encoded by the GBA1 gene, is a lysosomal hydrolase that converts glucosylceramide to glucose and ceramide. Mutations in GBA1 leading to GCase deficiency can cause lysosomal storage disorders. GCase deficiencies are implicated in several diseases, including Gaucher's disease and Parkinson's disease.
For Research Use Only. Not for use in diagnostic procedures.
Especificações
Cell PermeabilityCell-Permeant
ColorGreen
Excitation/Emission492/526
For Use With (Equipment)Flow Cytometer, High Content Imaging Instrument, Fluorescent Microscope, Confocal Microscope
Label or DyeFluorescein
Product TypeSubstrate
Quantity5 mg
Shipping ConditionRoom Temperature
Storage RequirementsFreezer (-5°C to -30°C)
SubstrateGCase Substrate
Detection MethodFluorescence
FormPowder
Substrate PropertiesChemical Substrate
Substrate TypeBeta-Glucosidase Substrate
Target EnzymeBeta-Glucosidase
Unit SizeEach
Conteúdo e armazenamento
Store in freezer (-5°C to -30°C).
Citações e referências (16)
Citações e referências
Abstract
The Parkinson's disease risk gene cathepsin B promotes fibrillar alpha-synuclein clearance, lysosomal function and glucocerebrosidase activity in dopaminergic neurons.
Journal:Molecular neurodegeneration
PubMed ID:39587654
Variants in the CTSB gene encoding the lysosomal hydrolase cathepsin B (catB) are associated with increased risk of Parkinson's disease (PD). However, neither the specific CTSB variants driving these associations nor the functional pathways that link catB to PD pathogenesis have been characterized. CatB activity contributes to lysosomal protein degradation
Development of screening strategies for the identification of paramylon-degrading enzymes.
Journal:Journal of industrial microbiology & biotechnology
PubMed ID:30806871
Enzymatic degradation of the β-1,3-glucan paramylon could enable the production of bioactive compounds for healthcare and renewable substrates for biofuels. However, few enzymes have been found to degrade paramylon efficiently and their enzymatic mechanisms remain poorly understood. Thus, the aim of this work was to find paramylon-degrading enzymes and ways
Dysregulation of mitochondria-lysosome contacts by GBA1 dysfunction in dopaminergic neuronal models of Parkinson's disease.
Journal:Nature communications
PubMed ID:33753743
Mitochondria-lysosome contacts are recently identified sites for mediating crosstalk between both organelles, but their role in normal and diseased human neurons remains unknown. In this study, we demonstrate that mitochondria-lysosome contacts can dynamically form in the soma, axons, and dendrites of human neurons, allowing for their bidirectional crosstalk. Parkinson's disease
Flow Cytometry Measurement of Glucocerebrosidase Activity in Human Monocytes.
Journal:Bio-protocol
PubMed ID:33659542
Glucocerebrosidase (GCase) is an important enzyme for the metabolism of glycolipids. GCase enzyme deficiency is implicated in human disease and the efficient measurement of GCase activity is important for evaluating the efficacy of therapeutics targeting this enzyme. Existing approaches to measure GCase activity include whole blood mass spectrometry-based assays, where
Tau accumulation in degradative organelles is associated to lysosomal stress.
Journal:Scientific reports
PubMed ID:37865674
Neurodegenerative disorders are characterized by the brain deposition of insoluble amyloidogenic proteins, such as α-synuclein or Tau, and the concomitant deterioration of cell functions such as the autophagy-lysosomal pathway (ALP). The ALP is involved in the degradation of intracellular macromolecules including protein aggregates. ALP dysfunction due to inherited defects in