Solarbio Tissue and Blood Alkaline Phosphatase (AKP/ALP) Kit za ispitivanje aktivnosti

Tissue and Blood Alkaline Phosphatase (AKP/ALP) Kit za ispitivanje aktivnosti

Bilješka: Uzmite dva ili tri različita uzorka za predviđanje prije testa.

Operativna oprema: Spektrofotometar

Catalog Number: BC2140

Veličina: 50T/24s

Komponente:

Ekstrakcijski reagens: Liquid 30 mL×1 bottle, storage at 4℃.

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

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

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

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

Opis proizvoda:

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.

Potrebni reagensi i oprema, ali nisu pruženi:

Spektrofotometar, stolna centrifuga, transferpettor, 1 ML staklena kiveta, led i destilirana voda.

Postupak:

Ja. Enzim preparation:

Dodati 1 mL Extraction reagent to 0.1 g tissue, grind thoroughly. Centrifuge at 4℃, 10000 RPM za 10 minuta. Take the supernatant on the ice for test. Blood sample can be detected directly. Dilute with Extraction reagent if concentrate is high.

Ii. Odlučnost postupak:

1. Predgrijan spektrofotometar za 30 minuta, Podesite valnu duljinu na 510 NM, Postavite brojač na nulu s destiliranim
2. Add reagents in 1.5 mL cuvette as the following:

Iii. AKP/ALP activity calculation:

• Koncentracija proteina:

Definicija jedinice: 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 Prot) = =[C×(A3-A4)÷(A1-A2)×Vs]÷(Cpr×Vs)÷T=0.167×(A3-A4)÷(A1-A2)÷Cpr

• Težina uzorka:

Definicija jedinice: 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 težina) = =[C×(A3-A4)÷(A1-A2)×Vs]÷(W ÷ ve × vs)÷T=0.167×(A3-A4)÷(A1-A2)÷W

• Krv:

Definicija jedinice: 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;

Vrpca: Ukupni volumen reakcije, 1020 μL=1.02 mL; Vs: Supernatant volumen, 0.02 ml;

VE: Valum ekstrakcije, 1 ml; W: Težina uzorka, g;

T: Vrijeme reakcije, 15 minuta;

CPR: Koncentracija proteina uzorka, mg/ml.

Bilješka:

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

Eksperimentalni primjer:

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 masa) = 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, Operacija se provodi prema koracima određivanja, 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, the enzyme activity is calculated as follows: 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.

Povezani proizvodi：

BC2020/BC2025 Acetylcholinesterase(AchE) Kit za ispitivanje aktivnosti

BC2130/BC2135 Acid Phosphatase(ACP) Kit za ispitivanje aktivnosti

BC0840/BC0845 Carboxylesterase(CarE) Kit za ispitivanje aktivnosti