肝醣測定試劑盒

筆記: 測試前取兩個或三個不同的樣本進行預測.

操作設備: Spectrophotometer/ microplate reader

目錄編號: BC0345

尺寸:100T/96S

成分:

Extract reagent: 100毫升×1, 4℃保存.

Regent I: 粉末×1, storage at 4℃.10 mg of glucose, 添加 1 mL of distilled water to dissolve it before use. Diluted with distilled water to 0.1 mg/mL glucose solution for standby, ready to use. 0.1 mg/mL glucose standard solution, 4℃保存.

Regent Ⅱ: 粉末×1, 4℃保存.

工作溶液: Pour 6 mL of distilled water into Reagent Ⅱ and slowly pour 24 mL of concentrated sulfuric acid. Dissolve and mix thoroughly before use. Unused reagents are valid at 4 °C for one week.

產品描述

Glycogen is a high molecular polysaccharide composed of glucose units. It is one of the main storage forms of sugar. It is mainly stored in the liver and muscle as backup energy, and is called liver glycogen and muscle glycogen, 分別. Glycogen can regulate blood glucose concentration. Glycogen can be synthesized in the liver when blood glucose rises. When blood sugar decreases, liver glycogen is broken down into glucose to supplement blood sugar. 所以, liver glycogen is important to maintain the relative balance of blood sugar. Muscle glycogen is a form of glycogen storage in muscles. When lots of blood sugar is consumed during strenuous exercise, muscle glycogen cannot be broken down directly into blood sugar. It must first be broken down to produce lactic acid, which is circulated to the liver with the blood, and transformed into liver glycogen through glycogen glucose.

Determination principle: anthrone method. Glycogen is extracted with strong alkaline extract, and the glycogen content is measured using an anthrone method under strongly acidic conditions.

需要但未提供的試劑和設備.

Spectrophotometer/ microplate reader, 桌上型離心機, 移液管, 微型玻璃比色皿/96孔板, mortar, concentrated sulfuric acid (硫酸) 和蒸餾水.

程式:

我. Sample extraction:

1. Cells or bacteria: 收集 5-10 million bacteria or cells into a centrifuge tube, discard the supernatant after centrifugation; add 0.75mL of extraction reagent to ultrasonically break bacteria or cells (力量 20% or 200W, ultrasonic 3s, 10s interval, 重複 30 次) ); Transfer to a 10mL tube, boil in a boiling water bath for 20min (close tightly to prevent water loss), shake the tube every 5 min to fully mix; take out the tube and cool, take up to 5mL with distilled water and mix. Centrifuge at 8000g and 25℃ for 10min, take the supernatant for testing.

2. 組織: Weigh 0.1~0.2g sample and put it in a 10 mL tube, 添加 0.75 mL of Extraction solution, boil in boiling water bath for 20 分分鐘 (close tightly to prevent water loss), shake the test tube every 5 min to fully mix. After all the tissue dissolved, take out the tube and cool down, then make up to 5 mL 加蒸餾水. Centrifuge at 8000g and 25℃ for 10 分分鐘, take the supernatant for testing.

二. 決心 程式:

1. Preheat the spectrophotometer/ microplate reader for 30 分分鐘, 調整波長至 620 奈米, 用蒸餾器調零
2. Sampling table (add the following regents in EPtube)

攪拌均勻, place in a boiling water bath for 10 分分鐘 (close tightly to prevent water loss), cool, and take 200 μL into micro glass cuvette/96-well plate to read the absorbance of the blank tube, standard tube, and measurement tube at 620 奈米, and record them as A1, A2, and A3. The blank tube and standard tube need only be tested once.

三、. 計算:

1. 樣品重量

Sorbitol (mg/g fresh weight) =(Cs×V1)×(A3-A1)÷(A2-A1)÷(W×V1÷V2)÷1.11

= 0.450×(A3-A1)÷（A2-A1)÷W

2. 蛋白質濃度

Sorbitol (毫克/毫克蛋白質) = (Cs×V1)×(A3-A1) ÷(A2-A1) ÷(V1×Cpr) ÷1.11

=0.09×(A3-A1) ÷（A2-A1) ÷心肺復甦

3. The number of bacteria or cells:

Sorbitol（mg/104 cell）= (Cs×V1)×(A3-A1)÷(A2-A1)÷(number of bacteria or cells×V1÷V2) ÷1.11

= 0.450×(A3-A1)÷（A2-A1)÷number of bacteria or cells

1.11: It is a constant that glucose content converted to glycogen content, That is, the color of 111 μg of glucose with anthrone reagent is equivalent to that of 100 μg of glycogen with anthrone reagent.

Cs: the concentration of standard, 0.1 mg/mL V1: 樣品量, 0.06 毫升;

V2: Total sample volume, 5 毫升;

心肺復甦: 樣品蛋白濃度, 毫克/毫升; 瓦: 樣品重量, G

Number of bacteria or cells: 104 as the unit, 萬

筆記:

If A is greater than 1.4, dilute the sample with distilled water and multiply it by the corresponding dilution factor in the calculation formula.

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Technical Specification:

檢測極限: 0.002 毫克/毫升

The linear range: 0.003125-0.25 毫克/毫升