GLUCAN DETECTION
Endpoint Assay: Reconstituted Glucatell ® reagent is used at a ratio of 1:1 and a volume of 50 μL/well (55 tests/vial). Performing the Test Kinetic Assay: The Glucatell ® /sample mixture is in- cubated at 37±1°C in a microplate reader. Software is used to construct the standard curve and calculate glucan concentrations. Endpoint Diazo Assay: The Glucatell ® /sample mix- ture is incubated at 37±1°C in a microplate heating block for the recommended time period. 50 μL each of the three diazo reagents are then added to the mixture. Software is used to construct the standard curve and calculate glucan concentrations. Reconstitution Glucatell ® reagent can be reconstituted differently depending on the assay you use. For endpoint assays, use 2.8 mL of only Pyrosol ® reconstitution buffer. For kinetic assays, combine 2.8 mL each of Pyrosol ® and Reagent Grade Water. Stability Store all reagents at 2–8°C in the dark. Once reconstituted, Glucatell ® reagent should be stored at 2–8°C and used within 2 hours. Alternatively, recon- stituted Glucatell ® reagent can be frozen at -20°C for 20 days, thawed once and used. The diazo reagents should be used the day they are prepared. Product Applications • Analyzing final products for (1 g 3)- b - D -Glucan • Investigating BET Out of Specification results • Qualifying raw materials • Monitoring cellulosic filter extractables • Monitoring fungal fermentation processes • Analyzing fermentation and cell culture media • Monitoring airborne glucan burden
Glucatell ® Kit – (1 g 3)- b - D -Glucan Detection
The Glucatell ® kit is specific for detection of (1 g 3)- b - D -Glucan (BG). BG is considered a contaminant in parenteral drugs, solutions, and medical devices due to BG’s bioactivity as an activator of innate immunity and its capacity to enhance bacterial endotoxin tests (literature references to the bioactivity of BG may be found at www.acciusa.com/pdfs/BG_Biol_Activity_Ref_ List_PR16023.pdf). The Glucatell assay is based upon a modification of the Limulus Amebocyte Lysate (LAL) pathway. Glucatell ® reagent is processed to eliminate Factor C, and is therefore specific for (1 g 3)- b - D -Glu- can. The reagent does not react with other polysaccha- rides, including beta-glucans with different glycosidic linkages. Glucatell ® is a chromogenic reagent that may be used to perform either kinetic or endpoint assays in microplate readers. Sensitivity The sensitivity for Glucatell ® assay is determined by the lowest standard concentration on the standard curve used for the assay. The maximum sensitivity for Glu- catell ® is 3.125 pg/mL when used with a microplate reader. Sample to Glucatell ® Ratio Kinetic Assay: Reconstituted Glucatell ® reagent is used at a ratio of 1:4 and a volume of 100 μL/well (55 tests/vial).
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