Description
system_update_altDatasheet
| Product Name: | PicoSense Dual-Sugar Intestinal Permeability Assay Kit |
| Product Code: | ARMA00352 |
| Product Size: | 100 Mannitol + 100 Lactulose Tests |
| Detection Principle: | Fluorometric |
| Excitation/Emission: | Ex 535 nm / Em 587 nm |
| Sample Type: | Urine or Blood |
The PicoSense assay measures intestinal permeability using two non-metabolizable sugars. Mannitol is readily absorbed transcellularly, serving as a marker for total absorptive surface area. Lactulose is normally excluded by healthy tight junctions but penetrates via paracellular pathways if the mucosal barrier is compromised. Lactulose is hydrolysed to fructose and galactose; galactose is oxidised by galactose dehydrogenase forming NADH. Mannitol is oxidised by mannitol dehydrogenase, also forming NADH. NADH converts resazurin to resorufin, producing a large increase in fluorescence. The ratio of lactulose to mannitol indicates intestinal permeability.
A. Mannitol Determination:
| Sample Collection: | Urine before and for 6 hours after ingestion of lactulose/mannitol cocktail; pool collected samples |
| Sample Preparation: | Mix 100 µl urine with 100 µl Sample Clean-up Mix; incubate 30 min at 37°C; centrifuge ~16,000 × g for 5 min |
| Standard Range: | 0, 0.4, 0.8, 1.2, 1.6, 2.0 nmol Mannitol |
| Sample Volume: | 10 µl sample diluted to 125 µl total with DI water; 5 µl per well in duplicate |
| Incubation: | 60 µl Mannitol Reaction Mix, 30 min at 37°C, then add 15 µl Enhancer |
| Measurement: | Fluorescence at Ex/Em 535/587 nm |
B. Lactulose Determination:
| Standard Range: | 0, 0.4, 0.8, 1.2, 1.6, 2.0 nmol Lactulose |
| Sample Volume: | 25 µl per well in duplicate |
| Hydrolysis: | Add 2 µl β-Galactosidase and 3 µl Lactulose Buffer; 30 min at 37°C |
| Fluorescence Development: | 60 µl Lactulose Reaction Mix, 30 min at 37°C, then add 15 µl Enhancer |
| Measurement: | Fluorescence at Ex/Em 535/587 nm |
| Unopened Kit Storage: | -20°C |
| Sample Clean-up Mix: | Add 10 ml of 50% glycerol; -20°C |
| Lactulose Buffer, Mannitol Buffer: | 4°C; Ready to use |
| β-Galactosidase, Lactulose Enzyme Mix: | Reconstitute with 220 µl respective buffer; keep on ice; -20°C storage |
| Mannitol Enzyme Mix: | Reconstitute with 220 µl Mannitol Buffer; keep on ice; -20°C storage |
| Resazurin: | -20°C |
| Enhancer: | 4°C |
| Lactulose & Mannitol Standards: | -20°C; Ready to use |
| Before Use: | Bring all components to room temperature; centrifuge vials briefly before opening |
