Solarbio -Formaldehyd -Dehydrogenase (Fdh) Aktivitätstest Kit (Tierproben)

Produktinformationen

Katalognummer: BC4980

Spezifikationen: 50T/48S

Kit-Komponenten

• Extraktionslösung: 60 ml x 1 Flasche, Lagern Sie bei 2-8 ° C.
• Reagenz eins: 35 ml x 1 Flasche, Lagern Sie bei 2-8 ° C.
• Reagenz zwei: Pulver x 2 Fläschchen, store at -20°C
• Reagenz drei: Pulver x 2 Fläschchen, Lagern Sie bei 2-8 ° C.
• Reagenz vier: 3 ml x 1 Flasche, Lagern Sie bei 2-8 ° C.

Lösungsvorbereitung

1. Reagenz II: Hinzufügen 0.25 mL distilled water before use. 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): Destilliertes Wasser (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. Reagenz III: Hinzufügen 1.5 mL distilled water before use. Gründlich mischen. Unused reagents should be stored at -20℃ for two weeks

Produktbeschreibung

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.

Anmerkungen:

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-Quarzküvette
• Mortar/homogenizer
• Ice and distilled water

Operation Steps:

ICH. Probenbehandlung (the amount of sample to be tested can be adjusted appropriately, and the specific ratio can be referenced)

• Gewebe: According to the ratio of animal tissue weight (G): Extraktionslösungsvolumen (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, Zentrifuge bei 8000 G, 4°C für 10 Protokoll; Nimm den Überstand (if the supernatant is not clear enough, it is recommended to repeat the above centrifugation steps) and place it on ice for testing.

II. Determination Steps

1. Preheat the UV spectrophotometer for at least 30 Protokoll, Passen Sie die Wellenlänge an 340 nm, und Null mit destilliertem Wasser.
2. Preheat Reagent One and Reagent Four at 37°C for 10 minutes before use.
3. In a 1 ml-Quarzküvette, hinzufügen 100 μl der Probe, 550 μL of Reagent One, 250 μL of Reagent Two working solution, 50 μL of Reagent Three, Und 50 μL of Reagent Four in sequence. Mix thoroughly immediately and determine the absorbance A1 at 340 NM für 20 Sekunden. 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 für 20 seconds and the absorbance A2 after 5 Protokoll. Calculate ΔA = A2 – A1.

III. Calculation of FDH Activity in Animal Tissue

1. Calculation based on sample protein concentration:

Einheitendefinition: 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.