Solarbio Formaldeide Deidrogenasi (FDH) Kit di analisi dell'attività (Campioni animali)

Product Information

Numero di catalogo: BC4980

Specifiche: 50T/48S

Kit Components

• Soluzione di estrazione: 60 mL x 1 bottiglia, store at 2-8°C
• Reagente uno: 35 mL x 1 bottiglia, store at 2-8°C
• Reagente due: Powder x 2 vials, store at -20°C
• Reagente tre: Powder x 2 vials, store at 2-8°C
• Reagente quattro: 3 mL x 1 bottiglia, store at 2-8°C

Solution Preparation

1. Reagente II: Aggiungere 0.25 ml di acqua distillata prima dell'uso. 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 (μL): Acqua distillata (μL) =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. Reagente III: Aggiungere 1.5 ml di acqua distillata prima dell'uso. Mescolare accuratamente. Unused reagents should be stored at -20℃ for two weeks

Descrizione del prodotto

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.

Appunti:

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 Cuvetta in quarzo da ml
• Mortar/homogenizer
• Ice and distilled water

Operation Steps:

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

• Tessuto: According to the ratio of animal tissue weight (G): extraction solution volume (ml) = 1:5~10 (it is recommended to weigh 0.1 g of animal tissue and add 1 mL of extraction solution), homogenize in an ice bath, centrifugare a 8000 G, 4°C per 10 minuti; prendi il surnatante (if the supernatant is not clear enough, it is recommended to repeat the above centrifugation steps) and place it on ice for testing.

II. Fasi di determinazione

1. Preheat the UV spectrophotometer for at least 30 minuti, regolare la lunghezza d'onda su 340 nm, e zero con acqua distillata.
2. Preheat Reagent One and Reagent Four at 37°C for 10 minutes before use.
3. In a 1 Cuvetta in quarzo da ml, aggiungere 100 μL of sample, 550 μL of Reagent One, 250 μL of Reagent Two working solution, 50 μL of Reagent Three, E 50 μL of Reagent Four in sequence. Mix thoroughly immediately and determine the absorbance A1 at 340 nm per 20 secondi. 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 nm per 20 seconds and the absorbance A2 after 5 minuti. Calculate ΔA = A2 – A1.

III. Calculation of FDH Activity in Animal Tissue

1. Calculation based on sample protein concentration:

Definizione di unità: 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.