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Specifications
- Assay Type Detection Kit
- Sample Types Serum, Plasma, Buffers, Tissue Culture Media
- Sensitivity 0.493 mg/dL
- Species Species Independent
- Assay Duration 30 Minutes
- Samples/Plate 41 in Duplicate
- Readout Colorimetric, 560nm
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Standard Curve
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Description
Assay Principle:
The Galactose Colorimetric Detection Kit quantitatively measures galactose levels in serum, plasma, buffers, and tissue culture media. The Galactose Colorimetric Detection Kit is a Detection Kit with a run time of 30 minutes. Please read the complete kit insert for more information before performing this assay.
Use our provided Galactose Standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate. Add the HRP, Substrate, and Galactose Oxidase solutions to each well, tapping the plate to ensure sufficient mixing of reagents. Then, incubate the mixture at room temperature for 30 minutes. The color-generating reaction occurs when galactose oxidase reacts with galactose in the standard or sample to produce hydrogen peroxide, which, in the presence of HRP, reacts with the colorless Substrate to produce a pink-colored product.
After the 30-minute incubation, use a plate reader to detect and record the generated signal at 560nm. Use the intensity and the standard curve to calculate the Galactose concentration in the samples.
Background:
Galactose is a hexose sugar that differs from glucose by the configuration of the hydroxyl group at the carbon-4 position. As an anomeric mixture of α-D-galactose and β-D-galactose, this monosaccharide exists abundantly in milk, dairy products, and many other food types, such as fruits and vegetables.
The Na+/glucose co-transporters SGLT1 and SGLT2 mediate galactose absorption in humans from food across the brush border membrane of the proximal jejunum and renal epithelium. Additionally, adults can produce up to 2 grams of galactose per day. In cells, β-D-galactose is epimerized to α-D-galactose and converted to galactose-1-phosphate (Gal-1-P). In the presence of galactose-1-phosphate uridylyltransferase, Gal-1-P reacts with UDP-glucose to form UDP-galactose and glucose-1-phosphate.
The glucose-1-phosphate produced can enter the glycolytic pathway or react with UTP in the presence of UDP-glucose pyrophosphorylase to form a new molecule of UDP-glucose. This enzyme pathway comprises the evolutionarily conserved Leloir pathway of galactose metabolism. If the flow of galactose through the Leloir pathway is perturbed either due to congenital deficiency of any of the enzymes mentioned above or an overwhelming presence of this hexose, toxicity syndromes (galactosemia) will be observed.
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Structure
