Glucose(Glu) Colorimetric Assay Kit(GOD-POD Method)

K085

48T(32S)/ 96T(80S)

Microplate reader(500-510 nm)

Universal

Quantitative detection of the glucose (Glu) content in Whole blood, serum, plasma and tissue samples.

2-8°C for 6 months

See kit label

For research use only.

Glucose oxidase can catalyze the oxidation of glucose to gluconic acid to produce hydrogen peroxide. In the presence of chromogenic oxygen receptors, peroxidase catalyzes hydrogen peroxide and oxidizes pigment sources to form colored substances.

1.Microplate reader (500-510 nm)

2.96-well plate

3.Precision Micropipettor and clean disposable tips

4.Clean EP tubes

5.Normal saline (0.9% NaCl)，Double distilled water

6.Benchtop centrifuge

7.Constant temperature water bath/incubator

Serum and plasma: detect directly.

whole blood: Take fresh blood and add it into the tube containing anticoagulant (heparin as anticoagulant, heparin concentration: 10-12.5 IU/mL blood), mix it upside down, take 0.1 mL and add 0.4 mL of double distilled water, mix it fully for 1 min, stand for 15 min. In this way, 5 times dissolved blood is prepared, which is clarified and transparent under light observation to be measured.

Before the formal test, 2-3 samples with significant differences should be selected and diluted into different concentrations for pre-test.

The recommended dilution factor for different samples is as follows (for reference only):

Note: The diluent is normal saline (0.9% NaCl).

1.Equilibrate all reagents to room temperature before use.

2.The preparation of enzyme working solution：

Mix Reagent 1 and Reagent 2 at the volume ratio of 1:1 before use, and store at 2-8° C away from light for 24 h.

3.The preparation of control working solution：

Mix normal saline and Reagent 2 at the volume ratio of 1:1 before use, and store at 2-8° C away from light for 24 h.

4.The preparation of standards at different concentrations

1.Standard well: Take 3 μL of standard solution with different concentration to the wells.

Sample well: Take 3 μL of sample to the wells.

Control well: Take 3 μL of sample to the wells.

2.Add 300 μL of enzyme working solution into the standard and sample well.

Add 300 μL of control working solution into the control well.

3.Cover the plate sealer and incubate at 37 °C for 15 min. Measure the OD value of each well with microplate reader at 505 nm.

Note: Set control wells for whole blood, hemolysis serum and plasma samples, but not for normal serum, plasma and tissue samples.

the standard curve: y=ax+b

Normal serum (plasma) sample: Glu content (mmol/L) = (ΔA505 - b) ÷ a × f

Whole blood and hemolysis sample: Glu content (mmol/L) = (ΔA’ - b) ÷ a × f

Note: y：OD_Standard – OD_Blank; x：Concentration of standard; a：The slope of the standard curve; b：The intercept of the standard curve; ΔA505：OD_Sample – OD_Blank; ΔA’：OD_Sample – OD_Control; f：Dilution factor of sample before tested.

1.This assay kit is for Research Use Only. Please read the instructions carefully and adjust the instruments before the experiments. Please follow the instructions strictly during the experiments. Protection methods must be taken by wearing lab coat and latex gloves.

2.The detection range of the kit is not equivalent to the concentration range of the substance to be measured in the sample. If the concentration of substance is not within the detection range exactly, an extra dilution or concentration should be taken for the sample. It is recommended to take a pre-test if your sample is not listed in the instruction book.

3.The experimental results are closely related to the situation of reagents, operations, environment and so on. Fine Test will guarantee the quality of the kits only, and NOT be responsible for the sample consumption caused by using the assay kits. It is better to calculate the possible usage of sample and reserve sufficient samples before use.