Solarbio甲醛去氫酶 (外傭) 活性測定試劑盒 (動物樣本)

產品資訊

目錄編號: BC4980

規格: 50T/48S

套件組件

• 萃取液: 60 毫升× 1 瓶子, 儲存於2-8°C
• 試劑一: 35 毫升× 1 瓶子, 儲存於2-8°C
• 試劑二: 粉x 2 小瓶, store at -20°C
• 試劑三: 粉x 2 小瓶, 儲存於2-8°C
• 試劑四: 3 毫升× 1 瓶子, 儲存於2-8°C

溶液製備

1. 試劑二: 添加 0.25 使用前毫升蒸餾水. 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 (微升): 蒸餾水 (微升) =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. 試劑三: 添加 1.5 使用前毫升蒸餾水. 充分混合. Unused reagents should be stored at -20℃ for two weeks

產品描述

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.

筆記:

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 mL quartz cuvette
• Mortar/homogenizer
• Ice and distilled water

Operation Steps:

我. 樣品處理 (the amount of sample to be tested can be adjusted appropriately, and the specific ratio can be referenced)

• 組織: According to the ratio of animal tissue weight (G): 萃取液體積 (毫升) = 1:5〜10 (建議稱重 0.1 g of animal tissue and add 1 mL of extraction solution), homogenize in an ice bath, 離心機在 8000 G, 4°C 為 10 分分鐘; 取上清液 (if the supernatant is not clear enough, it is recommended to repeat the above centrifugation steps) and place it on ice for testing.

二. Determination Steps

1. Preheat the UV spectrophotometer for at least 30 分分鐘, 調整波長至 340 奈米, 並用蒸餾水調零.
2. Preheat Reagent One and Reagent Four at 37°C for 10 minutes before use.
3. In a 1 mL quartz cuvette, 添加 100 µL 樣品, 550 μL of Reagent One, 250 μL of Reagent Two working solution, 50 μL of Reagent Three, 和 50 μL of Reagent Four in sequence. Mix thoroughly immediately and determine the absorbance A1 at 340 奈米對於 20 秒. 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 奈米對於 20 seconds and the absorbance A2 after 5 分分鐘. Calculate ΔA = A2 – A1.

三、. Calculation of FDH Activity in Animal Tissue

1. Calculation based on sample protein concentration:

單位定義: 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.