Jaringan Solarbio dan Fosfatase Alkalin Darah (AKP/ALP) Kit Uji Aktivitas

Fosfatase Alkalin Jaringan dan Darah (AKP/ALP) Kit Uji Aktivitas

Catatan: Ambil dua atau tiga sampel berbeda untuk prediksi sebelum pengujian.

Peralatan Operasi: Spektrofotometer

Nomor Katalog: BC2140

Ukuran: 50T/24S

Komponen:

Reagen ekstraksi: Cairan 30 botol ml × 1, penyimpanan pada suhu 4℃.

Reagen 1: Cairan 10 botol ml × 1, storage at 4℃ and protect from light.

Reagen 2: Cairan 10 botol ml × 1, storage at 4℃ and protect from light.

Reagen 3: Cairan 30 botol ml × 1, storage at 4℃ and protect from light. It cannot be used if it turns to blue-green.

Standar: Cairan 1 botol ml × 1, 10 μmol/mL phenol standard solution, penyimpanan pada suhu 4℃. Dilute with distilled water to 2.5 μmol/mL before use.

Deskripsi Produk:

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.

Reagen dan Peralatan Diperlukan tetapi Tidak Disediakan:

Spektrofotometer, mesin sentrifugal meja, TransferPettor, 1 mL kuvet kaca, es dan air suling.

Prosedur:

SAYA. Enzim persiapan:

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

II. Tekad prosedur:

1. Panaskan spektrofotometer untuk 30 menit, sesuaikan panjang gelombangnya 510 nm, Atur penghitung ke nol dengan suling
2. Add reagents in 1.5 mL cuvette as the following:

Nama reagen (μL)	Test tube A3	Control tube A4	Blank tube A2	Standard tube A1
Air sulingan			20
Solusi standar				20
Supernatan	20
Reagen 1	200	200	200	200
Reagen 2	200	200	200	200
Aduk rata, stay in 37℃ for 15 menit.
Reagen 3	600	600	600	600
Supernatan		20
Aduk rata, detect absorbance at 510 nm, record as A1, A2, A3, A4.

AKU AKU AKU. AKP/ALP activity calculation:

• Konsentrasi protein:

Definisi satuan: 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 keuntungan) =[C×(A3-A4)−(A1-A2)×Vs]−(Cpr×Vs)÷T=0.167×(A3-A4)−(A1-A2)−Cpr

• Berat sampel:

Definisi satuan: 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 (berat U/g) =[C×(A3-A4)−(A1-A2)×Vs]−(W÷Ve×Vs)÷T=0.167×(A3-A4)−(A1-A2)−W

• Darah:

Definisi satuan: 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: Konsentrasi standar, 2.5 mol/mL;

Tali: Volume reaksi total, 1020 μL=1.02 mL; Vs: Volume supernatan, 0.02 ml;

Ve: Volume ekstraksi, 1 ml; W: Berat sampel, G;

T: Waktu reaksi, 15 menit;

Cpr: Konsentrasi protein sampel, mg/mL.

Catatan:

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

Contoh eksperimental:

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 (massa U/g) = 0.167×(DI-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, Pengoperasiannya dilakukan sesuai dengan langkah-langkah penentuan, 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×(DI-AC) − (AS-AB) =0.167×(0.190-0.015)−(0.677-0.022)=0.0446U/mL.

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