Micro Total Carbohydrate Assay Kit

Catalogue No.: 
K139
Size: 
96T
Storage: 
The kit should be stored at 2-8℃ shading light for one year.
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  • SPECIFICATIONS
  • FIGURES
  • CONDITIONS
Catalogue No.

K139

Size

96T

Range

0.1-2mg/mL

Sensitivity

0.1mg/mL

Kit component

Reagents

96T

Storage

Hydrochloric Acid

120mL

2-8 °C

Sodium Hydroxide

120mL

2-8 °C

DNS Reagent

4.2mL

2-8 °C protected from light

Standard

Powder × 1 vial

2-8 °C

Storage

The kit should be stored at 2-8℃ shading light for one year.

Application

Detect the total Carbohydrate content in biological samples, including serum (or plasma), animal tissues, and plant tissues.

Principle of the Assay

Carbohydrate substances are one of the important components that make up the bodies of animals and plants, and they are also the main raw materials and storage substances for metabolism. Total Carbohydrate mainly refers to reducing sugars such as glucose, fructose, and lactose, as well as sucrose, maltose and possibly partially hydrolyzed starch that can be hydrolyzed into reducing monosaccharides under certain measurement conditions.

The detection principle of this kit is as follows: total Carbohydrate is hydrolyzed into reducing sugars. These reducing sugars react with DNS reagent under alkaline conditions to reduce DNS and generate amino compounds. These compounds appear red-brown in alkaline solution and have a characteristic absorption peak at 540 nm. Within a certain concentration range, the content of reducing sugars has a linear relationship with the absorbance at 540 nm. Based on the standard curve, the total Carbohydrate content in the sample can be calculated.

Additional Materials Required

Microplate reader or visible spectrophotometer (capable of measuring absorbance at 540nm), water bath, low-temperature centrifuge, 96-well plate or micro glass cuvette, adjustable pipette and pipette tips, homogenizer (for tissue samples), deionized water

Calculation

Note: The calculation formulas we provide for you include the derivation process formulas and the concise calculation formulas. Both are exactly the same. It is recommended to use the concise calculation formula in bold as the final calculation formula.

1. Drawing of standard curve

With the concentration of the standard solution as the y-axis and the ΔAStandard as the x-axis, draw the standard curve. Substitute ΔA into the standard curve formula to calculate y (mg/mL).

2. Calculation of total Carbohydrate content in the sample

(1) Calculated by protein concentration

Total sugar(mg/mg prot)=(y×V sample)÷(V sample×Cpr)×n=y÷Cpr×n

(2) Calculated by fresh weight of samples

Total sugar(mg/g)=(y×V sample)÷(W×V sample÷VS Sample Total)×n=10×y÷W×n

(3) Calculated by volume of Liquid samples

Total sugar(mg/mL)=y×V sample÷(V Liquid×V sample÷VL Sample Total)×n=10×y×n

Note:

V sample: Add sample volume, 0.03 mL;

VS Sample Total: Total volume of the sample, 10 mL;

Cpr: Sample protein concentration, mg/mL;

W: weight of sample, g;

V Liquid: Volume of the liquid sample, 0.1 mL;

VL Sample Total: Total volume of the liquid sample, 1 mL;

n: Dilution factor.

Notes

1.For tissue samples and cell samples, the results can be normalized between samples by measuring protein concentration. It is recommended to use the protein quantitative kit K001 (BCA method) from FineTest.

2.This kit is compatible with spectrophotometer detection. Please adjust the amount of test reagents according to the requirements of spectrophotometer detection in proportion.

3.It is recommended that the experimenters establish their own standard curves to improve the accuracy of the results. If they do not establish their own standard curves, they can use the typical standard curve formulas presented in the result display section for calculation.

4.This product is only limited to scientific research personnel.

5.Please pay attention to safety precautions and follow the operating procedures for laboratory reagents. For your safety and health, please wear a lab coat and use disposable gloves during the operation.

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