Соларбио формальдегиддегидрогеназа (ФДХ) Набор для анализа активности (Образцы животных)

Информация о товаре

Номер каталога: BC4980

Технические характеристики: 50Т/48С

Компоненты комплекта

• Решение для экстракции: 60 mL x 1 бутылка, хранить при 2-8°C
• Реагент Один: 35 mL x 1 бутылка, хранить при 2-8°C
• Реагент второй: Powder x 2 флаконы, store at -20°C
• Реагент Три: Powder x 2 флаконы, хранить при 2-8°C
• Реагент четвертый: 3 mL x 1 бутылка, хранить при 2-8°C

Solution Preparation

1. Реагент II: Добавлять 0.25 мл дистиллированной воды перед использованием. Unused reagents should be stored at -20℃ for two weeks.
2. Working solution of Reagent II: According to the amount required for the test, prepare the Working solution according to the ratio of Reagent II (мкл): Дистиллированная вода (мкл) =1:29, and prepare the reagents when it will be used. The Working solution of Reagent II should be used up on the same day if it is prepared on the same day.
3. Реагент III: Добавлять 1.5 мл дистиллированной воды перед использованием. Тщательно перемешать. Unused reagents should be stored at -20℃ for two weeks

Описание продукта

Formaldehyde dehydrogenase exists in most prokaryotes and all eukaryotes. It is an oxidoreductase that converts formaldehyde. Formaldehyde dehydrogenase can catalyze formaldehyde and NAD+ to produce NADH. The absorbance at 340 nm will increase. By measuring the change at 340nm, the activity of formaldehyde dehydrogenase can be calculated.

Примечания:

1. It is recommended to select 2-3 samples with expected large differences for pre-experiment before the experiment. If the sample absorbance is not within the measurement range, it is recommended to dilute or increase the sample volume for detection.

Required Instruments and Supplies:

• UV spectrophotometer
• Low-temperature centrifuge
• Constant temperature incubator/water bath
• Adjustable pipette
• 1 мл кварцевая кювета
• Mortar/homogenizer
• Ice and distilled water

Operation Steps:

я. Sample Treatment (the amount of sample to be tested can be adjusted appropriately, and the specific ratio can be referenced)

• Салфетка: According to the ratio of animal tissue weight (г): extraction solution volume (мл) "=" 1:5~10 (рекомендуется взвесить 0.1 g of animal tissue and add 1 mL of extraction solution), homogenize in an ice bath, центрифуга в 8000 г, 4°С для 10 минуты; возьмите супернатант (if the supernatant is not clear enough, it is recommended to repeat the above centrifugation steps) and place it on ice for testing.

II. Этапы определения

1. Preheat the UV spectrophotometer for at least 30 минуты, отрегулировать длину волны, чтобы 340 нм, и ноль дистиллированной водой.
2. Preheat Reagent One and Reagent Four at 37°C for 10 minutes before use.
3. В 1 мл кварцевая кювета, добавлять 100 μL of sample, 550 μL of Reagent One, 250 μL of Reagent Two working solution, 50 μL of Reagent Three, и 50 μL of Reagent Four in sequence. Mix thoroughly immediately and determine the absorbance A1 at 340 нм для 20 секунды. Place it in a 37°C water bath or incubator for 5 minutes to react, quickly remove it and wipe it dry, and determine the absorbance A2 at 5min20s. Record the absorbance A1 at 340 нм для 20 seconds and the absorbance A2 after 5 минуты. Calculate ΔA = A2 – А1.

III. Calculation of FDH Activity in Animal Tissue

1. Calculation based on sample protein concentration:

Определение единицы измерения: One unit of enzyme activity is defined as the amount of enzyme that catalyzes the production of 1 nmol of NADH per minute per mg of tissue protein.