Набор для анализа гликогена

Примечание: Перед тестированием возьмите два или три разных образца для прогнозирования..

Операционное оборудование: Spectrophotometer/ microplate reader

Номер каталога: BC0345

Размер:100Т/96С

Компоненты:

Extract reagent: 100мл×1, хранение при 4℃.

Регент I: Порошок×1, storage at 4℃.10 mg of glucose, добавлять 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, хранение при 4℃.

Regent Ⅱ: Порошок×1, хранение при 4℃.

Рабочее решение: 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.

Описание продукта

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, соответственно. 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. Поэтому, 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.

Требуемые реагенты и оборудование, которые не входят в комплект поставки.

Сpectrophotometer/ microplate reader, настольная центрифуга, пипетка для переноса, micro glass cuvette/96-well plate, mortar, concentrated sulfuric acid (H2SO4) и дистиллированная вода.

Процедура:

я. Sample extraction:

1. Cells or bacteria: Собирать 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 (власть 20% or 200W, ultrasonic 3s, 10s interval, repeat 30 раз) ); 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. Салфетка: Weigh 0.1~0.2g sample and put it in a 10 mL tube, добавлять 0.75 mL of Extraction solution, boil in boiling water bath for 20 мин (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 with distilled water. Centrifuge at 8000g and 25℃ for 10 мин, take the supernatant for testing.

II. Определение процедура:

1. Preheat the сpectrophotometer/ microplate reader for 30 мин, отрегулировать длину волны, чтобы 620 нм, set zero with distilled
2. Sampling table (add the following regents in EPtube)

Хорошо перемешать, place in a boiling water bath for 10 минуты (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 нм, and record them as A1, А2, and A3. The blank tube and standard tube need only be tested once.

III. Расчет:

1. Вес образца

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

= 0.450×(A3-A1)÷（A2-A1)÷Вт

2. Концентрация белка

Sorbitol (mg/mg prot) "=" (Cs×V1)×(A3-A1) ÷(A2-A1) ÷(V1×Cpr) ÷1.11

=0.09×(A3-A1) ÷（A2-A1) ÷Cpr

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: объем пробы, 0.06 мл;

V2: Total sample volume, 5 мл;

КПР: концентрация белка в образце, мг/мл; Вт: Вес образца, г

Number of bacteria or cells: 104 as the unit, ten thousand

Примечание:

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:

The detection limit: 0.002 мг/мл

The linear range: 0.003125-0.25 мг/мл