BioReagent BioReagent for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
Store at 2-8°C,Protected from light Ships Wet ice Check lot-specific COA for exact specifications.
SDS, COA, datasheet, and spec sheet available for download. Lot-specific COA accessible via lot number lookup.
Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
Reducing sugars (RS) are widely present in animals, plants, microorganisms, and cultured cells. Reducing sugars in plants primarily include glucose, fructose, and maltose. Among these, glucose and fructose are not only the main substrates for respiration but also serve as substrates for the further synthesis of sucrose, starch, and cellulose.
Detection Principle: In an alkaline solution, 3,5-dinitrosalicylic acid (DNS) can be reduced by reducing sugars to produce a brown-red-colored amino compound, which has a characteristic absorption peak at 540 nm. Within a certain concentration range, the RS content is linearly correlated with the absorbance at 540 nm. The RS content in the sample can be calculated based on a standard curve.
Detection Range: 0.05 - 0.6 mg/mL
Sensitivity: 0.025 mg/mL
Applicable Samples: Plant tissues, animal tissues, cells, bacteria, serum (plasma)
| R1501790 | Component | 48T | 96T | Storage |
| R1501790A | Extraction Buffer | 60 mL | 120 mL | 2-8℃ |
| R1501790B | DNS Reagent | 10 mL | 20 mL | 2-8℃. Store in the dark. |
| R1501790C | Standard | 1EA | 1EA | 2-8℃ |
Note: Before formal testing, it is recommended to perform a preliminary test with 2-3 samples expected to have significant differences.
User-Prepared Instruments and Reagents
Microplate reader or visible spectrophotometer (capable of measuring absorbance at 540 nm)
96-well plate or micro glass cuvettes, adjustable micropipettes and tips
Centrifuge, water bath
Deionized water
Homogenizer (for tissue samples)
Experimental Procedure
1. Reagent Preparation
| Reagent Name | Reagent Preparation | Notes |
| Extraction Buffer | Ready-to-use; Equilibrate to room temperature before use. | Store at 4°C. Slightly irritating. Use appropriate personal protective equipment. |
| DNS Reagent | Ready-to-use; Equilibrate to room temperature before use. | Store at 4°C protected from light. Slightly irritating. Use appropriate personal protective equipment. |
| Standard | Before use, add 1 mL of deionized water to dissolve, preparing a 10 mg/mL stock standard solution. | Can be stored at 4°C for 2 weeks. |
2. Standard Curve Setup
Dilute the 10 mg/mL standard stock solution with deionized water to concentrations of 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, and 0.05 mg/mL.
| Tube | Volume of 10 mg/mL Standard (μL) | Volume of Deionized Water (μL) | Concentration (mg/mL) |
| Std.1 | 60 | 940 | 0.6 |
| Std.2 | 50 | 950 | 0.5 |
| Std.3 | 40 | 960 | 0.4 |
| Std.4 | 30 | 970 | 0.3 |
| Std.5 | 20 | 980 | 0.2 |
| Std.6 | 10 | 990 | 0.1 |
| Std.7 | 5 | 995 | 0.05 |
Note: The standard curve must be generated with each experiment. Diluted standard solutions are unstable and must be used within 4 hours.
3. Sample Preparation
3.1 Plant or Animal Tissue Samples
Weigh approximately 0.1 g of tissue. Add 1 mL of Extraction Buffer and homogenize in an ice bath. Transfer the homogenate to a capped centrifuge tube (to prevent evaporation during heating). Incubate in an 80°C water bath for 40 minutes, vortexing every 5 minutes. Centrifuge at 8,000 g, 25°C for 10 minutes. Collect the supernatant for assay.
3.2 Bacteria or Cells
Collect bacteria or cells into a centrifuge tube; discard the supernatant. Add 1 mL of Extraction Buffer per 5 million bacteria/cells. Sonicate in an ice bath for 5 minutes (power 20%, pulse 3s on, 10s off, repeat 30 times). Transfer to a capped centrifuge tube (to prevent evaporation during heating). Incubate in an 80°C water bath for 40 minutes, vortexing every 5 minutes. Centrifuge at 8,000 g, 25°C for 10 minutes. Collect the supernatant for assay.
3.3 Serum (Plasma) Samples
Take 0.1 mL of serum (plasma) and add 0.9 mL of Extraction Buffer; mix thoroughly. Transfer to a capped centrifuge tube (to prevent evaporation during heating). Incubate in an 80°C water bath for 40 minutes, vortexing every 5 minutes. Centrifuge at 8,000 g, 25°C for 10 minutes. Collect the supernatant for assay.
Note:
If protein concentration measurement is required, Aladdin's BCA Protein Quantification Kit (B665595) or Ready-to-Use BCA Protein Quantification Kit (R1491648) is recommended. The Extraction Buffer contains components that denature proteins. If calculating based on protein concentration, protein needs to be re-extracted separately for measurement.
4. Assay Steps
4.1 Preheat the microplate reader or visible spectrophotometer for at least 30 minutes. Set the wavelength to 540 nm. For spectrophotometers, zero the instrument with deionized water.
4.2 Assay Procedure:
| Reagent | Blank Tube (μL) | Standard Tube (μL) | Test Tube (μL) | Control Tube (μL) |
| Sample | 0 | 0 | 175 | 175 |
| Standard (various conc.) | 0 | 175 | 0 | 0 |
| Deionized Water | 175 | 0 | 0 | 125 |
| DNS Reagent | 125 | 125 | 125 | 0 |
Mix well. Heat in a boiling water bath for 5 minutes (cap tightly to prevent evaporation). Remove and immediately cool to room temperature. Transfer 200 μL to a 96-well plate or micro glass cuvette. Measure the absorbance at 540 nm. Calculate ΔA test = A test - A control , ΔA standard = A standard - A blank .
Note:
The Blank and Standard tubes only need to be set up 1-2 times.
It is recommended to perform a preliminary test with 2-3 samples expected to have significant differences before the formal experiment. If ΔA <sub> test </sub> is less than 0.04, consider increasing the sample volume appropriately. If ΔA <sub> test </sub> is greater than the ΔA <sub> standard </sub> of the 0.6 mg/mL standard, further dilute the sample with Extraction Buffer (multiply the result by the dilution factor) or reduce the amount of sample used for extraction.
5. Calculation of Results
Note: We provide both the derived formula and a simplified formula. They are equivalent. It is recommended to use the simplified formula in bold for final calculation.
5.1 Standard Curve Plotting
Plot the standard concentration (y-axis) against ΔA standard (x-axis) to generate the standard curve. Substitute ΔA test into the standard curve equation to calculate y (mg/mL).
5.2 Sample Reducing Sugar Content Calculation
(1) Based on Sample Weight
Reducing Sugar (μg/g) = 1000 × y × V<sub>extraction</sub> ÷ W × n = 1000 × y / W × n
(2) Based on Sample Protein Concentration
Reducing Sugar (μg/mg prot) =1000 × y × Vextraction ÷ (Vextraction × Cpr) × n=1000 × y / Cpr × n
(3) Based on Bacterial or Cell Count
Reducing Sugar (μg/10⁴) =1000 × y × V<sub>extraction</sub> ÷ 500 × n = 2 × y × n
(4) Based on Serum (Plasma) Volume
Reducing Sugar (μg/mL) = 1000 × y × Vextraction ÷ Vliquid × n = 10000 × y × n
Parameter Definitions:
1000: Unit conversion factor (1 mg/mL = 1000 μg/mL)
V extraction : Volume of Extraction Buffer added (1 mL)
V liquid : Volume of serum (plasma) added (0.1 mL)
Cpr: Sample protein concentration (mg/mL)
W: Sample weight (g)
500: Total number of bacteria or cells (5 million)
n: Dilution factor
6. Representative Results
Typical Standard Curve: y = 0.2243x + 0.0545, R² = 0.9957
Precautions
This product is for research use only. Not for use in clinical diagnosis. For your safety and health, please wear lab coats and disposable gloves during operation.
Comprehensive hazard, handling, storage, and regulatory compliance document.
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