Solarbio Tissue and Blood Alkaline Phosphatase (AKP/ALP) Aktivitätstest-Kit

Tissue and Blood Alkaline Phosphatase (AKP/ALP) Aktivitätstest-Kit

Notiz: Nehmen Sie vor dem Test zwei oder drei verschiedene Proben zur Vorhersage.

Betriebsausrüstung: Spektrophotometer

Katalognummer: BC2140

Größe: 50T/24s

Komponenten:

Extraktionsreagenz: Flüssig 30 mL×1 bottle, storage at 4℃.

Reagens 1: Flüssig 10 mL×1 bottle, storage at 4℃ and protect from light.

Reagens 2: Flüssig 10 mL×1 bottle, storage at 4℃ and protect from light.

Reagens 3: Flüssig 30 mL×1 bottle, storage at 4℃ and protect from light. It cannot be used if it turns to blue-green.

Standard: Flüssig 1 mL×1 bottle, 10 μmol/mL phenol standard solution, storage at 4℃. Dilute with distilled water to 2.5 μmol/mL before use.

Produktbeschreibung:

AKP/ALP is a zinc-containing glycoprotease, which hydrolysis various natural and synthetic phospholipid monoester compounds in alkaline condition. AKP / ALP are widely distributed in human organs, mainly in liver.

In alkaline condition, AKP/ALP catalyzes hydrolysis of disodium phenyl phosphate to phenol, and the phenol reacts with 4-Aminoantipyrine and potassium ferricyanide to form red quinone derivative, which can be detected absorbance at 510 nm. AKP/ALP activity can be calculated by measuring the absorbance increase rate at 510 nm.

Erforderliche, aber nicht bereitgestellte Reagenzien und Ausrüstung:

Spektrophotometer, Tischzentrifuge, transferpettor, 1 ML Glass Cuvette, Eis und destilliertes Wasser.

Verfahren:

ICH. Enzym Vorbereitung:

Hinzufügen 1 mL Extraction reagent to 0.1 g tissue, grind thoroughly. Centrifuge at 4℃, 10000 U/min für 10 Protokoll. Take the supernatant on the ice for test. Blood sample can be detected directly. Dilute with Extraction reagent if concentrate is high.

II. Bestimmung Verfahren:

1. Spektrophotometer vorheizen 30 Protokoll, Passen Sie die Wellenlänge an 510 nm, Stellen Sie den Zähler mit destilliert auf Null ein
2. Add reagents in 1.5 mL cuvette as the following:

III. AKP/ALP activity calculation:

• Proteinkonzentration:

Einheitendefinition: One unit of enzyme activity is defined as the amount of enzymes catalyzes the generation of 1 μmol phenol in the reaction system per minute at 37℃ every mg protein.

AKP/ALP (U/mg-Schutz) =[C×(A3-A4)÷(A1-A2)×Vs]÷(CPR × vs)÷T=0.167×(A3-A4)÷(A1-A2)÷ CPR

• Probengewicht:

Einheitendefinition: One unit of enzyme activity is defined as the amount of enzymes catalyzes the generation of 1 μmol phenol in the reaction system per minute at 37℃ every g sample.

AKP/ALP (U/g-Gewicht) =[C×(A3-A4)÷(A1-A2)×Vs]÷(W÷Ve×Vs)÷T=0.167×(A3-A4)÷(A1-A2)÷ w

• Blut:

Einheitendefinition: One unit of enzyme activity is defined as the amount of enzymes catalyzes the generation of 1 μmol phenol in the reaction system per minute at 37℃ every mL serum.

AKP/ALP (U/ml) =[C×(A3-A4)÷(A1-A2)×Vs]÷Vs÷T=0.167×(A3-A4)÷(A1-A2)

C: Standard concentration, 2.5 μmol/ml;

Seil: Gesamtreaktionsvolumen, 1020 μL=1.02 mL; Vs: Supernatant volume, 0.02 ml;

Ve: Extraktionsvolumen, 1 ml; W: Probengewicht, G;

T: Reaktionszeit, 15 Protokoll;

Cpr: Proteinkonzentration der Probe, mg/ml.

Notiz:

1. Reagens 1, Reagens 2 and Reagent 3 should be protected from
2. Reagens 3 cannot be used if it has changed toblue-green.
3. Mix thoroughly quickly after adding Reagent 3 to avoid incomplete

Experimentelles Beispiel:

1. Take 0.1g of mouse pancreas and add 1 mL of Extract solution for homogenate. After taking the supernatant, operate according to the determination steps. Calculate AT = 0.324, Ac = 0.037, AB = 0.022, AS = 0.677. Calculate the enzyme activity according to the sample mass: AKP/ALP enzyme activity (U/g-Masse) = 0.167×(AT-AC)÷(AS-AB) ÷ W =0.167×(0.324-0.037)÷(0.677-0.022)÷0.1=0.732 U/g
2. After taking the rabbit serum, Die Operation wird gemäß den Bestimmungsschritten durchgeführt, and the enzyme activity is calculated as follows: AT = 0.190, Ac = 0.015, AB = 0.022, AS = 0.677. According to the blood volume, Die Enzymaktivität wird wie folgt berechnet: AKP/ALP enzyme activity (U/ml) = 0.167×(AT-AC) ÷ (AS-AB) =0.167×(0.190-0.015)÷(0.677-0.022)=0.0446U/mL.

Verwandte Produkte：

BC2020/BC2025 Acetylcholinesterase(AchE) Aktivitätstest-Kit

BC2130/BC2135 Acid Phosphatase(AKP) Aktivitätstest-Kit

BC0840/BC0845 Carboxylesterase(CarE) Aktivitätstest-Kit