Galactose Colorimetric/Fluorometric Assay Kit Kit
- Detection Method
- Fluorometric, Colorimetric
- Biochemical Assay (BCA)
- Sample Type
- Cell Culture Supernatant, Plasma, Serum, Tissue Samples
- Galactose Assay Kit provides a simple, convenient means for direct measurement of galactose levels in various biological samples (serum, plasma, other body fluids, food, growth media, etc.). In the assay kit, Galactose is specifically oxidized generating a product that produces color (O.D 570 nm) and fluorescence (Ex/Em 535/587 nm). Liquid samples can be tested directly without purification. The assay is fast, convenient and sensitive. It can also be used as a high throughput assay.
- Galactose Assay Kit: High-Throughput Colorimetric & Fluorometric Assay to Measure Galactose levels from various Biological Samples such as Serum, Plasma, other Body Fluids, Food, Growth media, etc. within 40 min. Rapid, Convenient & Sensitive.
Galactose Assay Buffer
Galactose Probe (DMSO solution)
Galactose Enzyme Mix (Lyophilized)
Galactose Standard (100 nmol/μl)
Further details regarding sample type: Cell and tissue culture supernatants, urine, plasma, serum, as well as many other biological fluids
- Assay Time
- < 1 h
1. Standard Curve Preparation: For the colorimetric assay, dilute the Galactose Standard to 1 nM/µL by adding 10 µL of the 100 nM/µL Galactose Standard to 990 µL of Galactose Assay Buffer and mix well. Add 0, 2, 4, 6, 8, 10 µL into a series of wells. Adjust the volume to 50 µL/well with Galactose Assay Buffer to generate 0, 2, 4, 6, 8, 10 nM/well of Galactose Standard. For the fluorometric assay, dilute the Galactose Standard solution to 0.1 nM/µL by adding 10 µL of the Galactose Standard to 990 µL of Galactose Assay Buffer and mix well. Then take 20 µL into 180 µL of Galactose Assay Buffer and mix well. Add 0, 2, 4, 6, 8, 10 µL into each well individually. Adjust volume to 50 µL/well with Galactose Assay Buffer to generate 0, 0.2, 0.4, 0.6, 0.8, 1.0 nM/well of the Galactose Standard.
2. Sample Preparation: Liquid samples can be directly added into the plate, then adjust to a total volume of 50 µL with Galactose Assay Buffer. For unknown samples, we suggest testing several doses of sample to ensure the readings are within the standard curve linear range.
3. Galactose Reaction Mix: Mix enough reagent for the number of assays to be performed. For each well, prepare a total 50 µL Reaction Mix containing: 44 µL Galactose Assay Buffer 2 µL Galactose Probe* 2 µL Galactose Enzyme Mix 2 µL HRP * Note: The fluorometric assay is approx. 10 fold more sensitive than the colorimetric assay. Use 0.4 µL probe in each reaction in the fluorometric assay to decrease background fluorescence significantly.
4. Mix well. Add 50 µL of the Reaction Mix to each well containing the Galactose Standard and test samples. Mix well.
5. Incubate the reaction for 30 minutes at 37 °C, protect from light.
6. Measure O.D. 570nm for the colorimetric assay or Ex/Em = 535/590 nm for the fluorometric assay in a microplate reader.
- Calculation of Results
Calculations: Correct background by subtracting the value of the 0 galactose control from all sample readings. Plot standard curve galactose amount (nmol) vs OD 570nm. Apply sample readings to the standard curve. Galactose concentration: C = Ga/Sv nM/µL or mM Where Ga: Galactose amount in the sample wells (in nmol). Sv: Sample volume added into the sample wells (in µL). Galactose Molecular Weight: 180.16.
- For Research Use only
- -20 °C
- Expiry Date
- 12 months
Sela, Garrido, Lerno, Wu, Tan, Eom, Joachimiak, Lebrilla, Mills: "Bifidobacterium longum subsp. infantis ATCC 15697 α-fucosidases are active on fucosylated human milk oligosaccharides." in: Applied and environmental microbiology, Vol. 78, Issue 3, pp. 795-803, 2012 (PubMed).
- Sela, Garrido, Lerno, Wu, Tan, Eom, Joachimiak, Lebrilla, Mills: "Bifidobacterium longum subsp. infantis ATCC 15697 α-fucosidases are active on fucosylated human milk oligosaccharides." in: Applied and environmental microbiology, Vol. 78, Issue 3, pp. 795-803, 2012 (PubMed).
- Target Type
- Galactose (C₆H₁₂O₆ FW: 180.16) is one the major naturally occurring sugars, involved in many biological processes.