Galactose Assay Kit
BCA
Cell Culture Supernatant, Milk, Plasma, Saliva, Serum, Urine
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(1 reference)-
- 抗原
- Galactose
- 应用范围
- Biochemical Assay (BCA)
- 样品类型
- Cell Culture Supernatant, Milk, Plasma, Saliva, Serum, Urine
- 特异性
- 10 μM
- 产品特性
- Use as little as 20 µL samples. Linear detection range in 96-well plate: 10 to 1000 µM galactose for colorimetric assays and 10 to 100 µM for fluorimetric assays.
- 组件
- Assay Buffer: 10 mL. Enzyme Mix: 120 µL. Dye Reagent: 120 µL. Standard: 1 mL 10 mM Galactose.
- 试剂未包括
- Pipetting devices, centrifuge tubes, clear flat-bottom 96-well plates, optical density plate reader, black 96-well plates and fluorescence plate reader.
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- 应用备注
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Direct Assays: galactose in serum, plasma, urine, saliva, milk, culture medium and other biological samples.
Drug Discovery/Pharmacology: effects of drugs on galactose metabolism.
Food and Beverages: galactose in food and beverages products. - 实验流程
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Note: (1) glycerol and SH-containing reagents (e.g. µMercaptoethanol, dithiothreitol) are known to interfere in this assay and should be avoided in sample preparation. (2) This assay is based on a kinetic reaction. To ensure identical incubation time, addition of Working Reagent to standard and samples should be quick and mixing should be brief but thorough. Use of a multi-channel pipettor is recommended.
1. Equilibrate all components to room temperature. During experiment, keep thawed Enzyme Mix in a refrigerator or on ice.
2. Standards and samples: prepare 400 µL 1000 µMStandard by mixing 40 µL 10 mM standard with 360 µL dH2O. Dilute standard in dH2O. Transfer 20 µL standards and 20 µL samples into separate wells of a clear flat-bottom 96-well plate.
3. Reaction. For each reaction well, mix 85 µL Assay Buffer, 1 µL Enzyme Mix (vortex briefly before pipetting), and 1 µL Dye Reagent in a clean tube. Transfer 80 µL Working Reagent into each reaction well. Tap plate to mix. Incubate 20 min at room temperature.
4. Read optical density at 570nm (550-585nm). - 样品制备
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Serum and plasma samples can be assayed directly. Milk samples should be cleared by mixing 600 µL milk with 100 µL 6 N HCl. Centrifuge 5 min at 14,000 rpm. Transfer 300 µL supernatant into a clean tube and neutralize with 50 µL 6 N NaOH. The neutralized supernatant is ready for assay (dilution factor n = 1.36)
- 结果分析
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Subtract blank value (water, #8) from the standard values and plot the OD or RFU against standard concentrations.
Conversions: 1 mM galactose equals 18 mg/dL, 0.018% or 180 ppm. - 限制
- 仅限研究用
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- 储存条件
- -20 °C
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: "An investigation into the relationship between metabolic responses and energy regulation in antibody-producing cell." in: Journal of microbiology and biotechnology, Vol. 23, Issue 11, pp. 1586-97, (2013) (PubMed).
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: "An investigation into the relationship between metabolic responses and energy regulation in antibody-producing cell." in: Journal of microbiology and biotechnology, Vol. 23, Issue 11, pp. 1586-97, (2013) (PubMed).
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- 抗原
- Galactose
- 背景
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Quantitative determination of galactose by colorimetric (570nm) or fluorimetric (530nm/590nm) methods.
Procedure: 20 min.
Galactose (C6H12O6) is a monosaccharide that is found in dairy products, sugar beets, gums and mucilages. It is also synthesized in mammals, where it forms part of glycolipids and glycoproteins in several tissues. It forms the disaccharide lactose when combined with glucose. Simple, direct and high-throughput assays for galactose determination find wide applications. This assay uses specific enzyme-coupled reactions to form a colored product. The color intensity at 570nm or fluorescence intensity at 530nm/585nm is directly proportional to the galactose concentration in the sample.
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