Plant Dehydrogenase (PDHA) Aktivitätstest-Kit
Notiz: Nehmen Sie vor dem Test zwei oder drei verschiedene Proben zur Vorhersage.
Betriebsausrüstung: Microplate Reader/ Spectrophotometer
Katze nein: BC3125
Größe:100T/48S
Komponenten:
Reagenz I: Pulver×2. Lagerung bei 4℃. Dissolve one bottle of powder in water to make 50 mL before use, prepare the solution when it will be use. Store at 4℃ and protect from light after prepared.
Reagenz II: Flüssig 100 mL×2. Lagerung bei 4℃.
Reagenz III: Ethyl acetate, required but not provided.
Produktbeschreibung:
The activity of plant dehydrogenase (PDHA) is largely reflecting the active state of the organism, which can directly indicate the ability of biological cells to degrade its matrix.
The hydrogen acceptor 2,3,5-triphenyl tetrazolium chloride (TTC) generates red triphenylformazone (TFF) after receiving hydrogen during cell respiration. TFF has a characteristic absorption peak at 485 nm, the PDHA activity can quantified by measuring the absorbance at 485 nm.
Erforderliche, aber nicht bereitgestellte Reagenzien und Ausrüstung:
Microplate Reader or spectrophotometer, Wasserbad, Tischzentrifuge, Wasserbad, Pipette, Mikroglasküvette/96-Well-Platte mit flachem Boden (non-polystyrene/polypropylene material), Mörtel/Homogenisator, ethyl acetate (express delivery is not allowed), Eis und destilliertes Wasser.
Verfahren:
ICH. Complex extraction:
Sammeln 0.1 G Gewebe, waschen 3 oder 4 times with double steam water, blot dry with filter paper and set aside.
II. Bestimmung Verfahren:
- Preheat spectrophotometer/ microplate reader for 30 Protokoll, Passen Sie die Wellenlänge an 485 nm, set zero with ethyl acetate.
- Fügen Sie die folgenden Reagenzien hinzu 5 mL EP tubes:
| Reagens | Contrast tube (AC) | Reagenzglas (BEI) |
| Probe (G) | 0.1 | 0.1 |
| Reagenz I (ml) | – | 1 |
| Reagenz II (ml) | 2 | 1 |
| Mix thoroughly and stand in dark for 3 hours at 37℃, ice bath for 5 minutes immediately after take out. Discard the filtrate, blot dry the sample with filter paper, place in mortar / homogenizer. | ||
| Reagenz III (ml) | 1 | 1 |
III. Berechnung:
A. micro glass cuvette (light path of the cuvette, 1 cm)
Einheitendefinition: One unit of enzyme activity is defined as the amount of enzyme increases the absorbance of every 0.01 for per hour every mg tissue protein in the reaction system.
PDHA Activity(U/g/h) =ΔA÷0.01÷T÷W=33×ΔA÷W
B. 96 Brunnenplatte (light path of the cuvette, 0.6 cm)
Einheitendefinition: One unit of enzyme activity is defined as the amount of 1 mg of tissue increases the absorbance of every 0.005 for per hour in the reaction system.
PDHA Activity(U/g/h) =ΔA÷0.005÷T÷W=66.7×ΔA÷W
T: Reaktionszeit (H), 3 Std.; W: Probengewicht, G.
Notiz
- After prepared, Reagent I should store at 4℃, protect from light and used within one week. If it turns red, it cannot be used.
- Reagent III is volatile and toxic. For your health, please wear lab clothes, masks, and latex gloves.
- Ice bath to stop the reaction immediately after the dark reaction was completed. Discard the filtrate, blot dry the sample with filter paper as much as possible.
- Nehmen Sie vor dem Test zwei oder drei verschiedene Proben zur Vorhersage. Dilute the supernatant if the absorbance is higher, multiply dilute times in the formula.
- If test with a 96 gut flacher Teller, polystyrene, or polypropylene material 96 well flat-bottom plate is not recommended.
Experimentelles Beispiel:
- Weigh 1g aloe leaves, wash them with double distilled water for 3–4 times, absorb the water with filter paper, operate according to the determination steps, measure and calculate with 96 Brunnenplatte, ΔA=AT-AC= 0.244-0.146 = 0.098, calculate the enzyme activity.
Dehydrogenase activity (U/g-Masse) = 66.7×ΔA÷W = 6.5366 U/g-Masse.
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BC2030/BC2035 Isocitrate Lyase (ICL) Aktivitätstest-Kit
BC3170/BC3175 Acetokinase (ACK) Aktivitätstest-Kit
BC2010/BC2015 Glycollic Oxidase Activity Assay Kit
Rezensionen
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