The Real Time Blues™ (RTB) - Lactose/D-galactose detection assay
Optimized for glycobiology applications
The Real Time Blues™ (RTB) - Lactose/D-galactose detection assay can be used to quantify the lactose or D-galactose concentration in an unknown sample. In this assay, lactose is hydrolysed by β-galactosidase to produce total D-glucose and D-galactose. Subsequently, galactose oxidase converts the D-galactose to D-galacto-hexodialdose and hydrogen peroxide. Finally, horseradish peroxidase is used to convert our RTB reagent and the resulting hydrogen peroxide into a stable blue product that can be measured optically. The amount of lactose is then calculated by subtracting the free D-galactose detection from the total D-galactose detection (sample with and without β-galactosidase hydrolysis).
What's Included
This kit provides enough reagents for conducting 100 assays of lactose or D-galactose in a 96-well optical plate using sample volumes of 100 µL per well. In this format as little as 2 nmoles of D-galactose or 4 nmoles of lactose can be quantified.
Principle | Specifications | Technical Info |
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 | > Quantitative colorimetric | > Robust reagent |
> Real-time or Endpoint | > 2 mg/L lactose detection |
> Broad range linearity | > 1 mg/L D-galactose detection |
> Stable coloration | |
> pH: 3.0 to 9.0 | |
| |
References
- Holland V.R., Saunders B.C., Rose F.L., Walpole A.L..”A safer substitute for benzidine in the detection of blood”. Tetrahedron 30: 3299 (1974).
- Josephy, P.D., Eling, T., and Mason, R.P. “The Horseradish Peroxidase-catalyzed Oxidation of 3,5,3’,5’-tetramethylbenzidine. Free radical and charge-transfer complex intermediates.” J. Biol. Chem. 257, 3669-3675 (1982).
- Marquez, L.A. and Dunford, H.B. Mechanism of the oxidation of 3,5,3’,5’-tetramethylbenzidine by myeloperxidase determined by transient-and steady-state kinetics. Biochemistry 36, 9349-9355 (1997).
- American Association of Cereal Chemists. 1983. Approved Methods of the AACC, 8th edition. Methods 80-68, 80-31 and 80-50. The Association; St. Paul, MN.