간 리파아제 (HL) 활동 분석 키트

메모: 테스트 전에 예측을 위해 2~3개의 서로 다른 샘플을 채취합니다..

운영장비: 분광 광도계

고양이 번호: BC2380

크기:50T/24S

구성요소:

시약 I: 100 밀리리터×1. 4℃에서 보관.

시약 II: 3 밀리리터×1. 4℃에서 보관.

시약 III: 가루×1. 4℃에서 보관. 사용하기 전에, 추가하다 30 증류수 1mL, fully dissolve.

시약 IV: 가루×2. -20℃에서 보관. 사용하기 전에, 추가하다 2 mL of distilled water to the one, fully dissolve. The dissolved reagent can be stored at -20 °C after repacking. 반복적 인 동결-해동 사이클을 피하십시오;

기준: 가루×1. 사용하기 전에, 6.94 mL of 아세톤 is added to prepare a 10 μmol/mL α-naphthol

표준 용액, which was fully dissolved before use.

제품 설명:

간 리파아제 (HL) is a lipolytic enzyme synthesized in liver parenchymal cells. It is present on the surface of the liver sinusoidal endothelial cells and the surface of the hepatocyte microvilli in the sinusoidal space, and can hydrolyze various lipoproteins. The triglycerides (TG) and phospholipids (PL) in the medium change the size and density of various lipoprotein particles. When the HL and its activity in the plasma increasing, it can lead to low density lipoprotein (LDL) levels in the plasma, increase and accelerate the occurrence and development of atherosclerosis.

HL hydrolyzes α-naphthyl acetate to produce α-naphthol, which can form a purple-red azo compound with fast blue B salt. It has a characteristic absorption peak at 595 nm, and its color depth is positively correlated with liver esterase activity within a certain range.

필요하지만 제공되지 않는 시약 및 장비:

분광 광도계, 욕조, 균형, 원심분리기, adjustable transferpettor, 1 mL 유리 큐벳, 모르타르/균질화기, ultrasonic crusher, 얼음과 증류수.

절차:

나. 효소 추출

1. 조직

According to the tissue mass (g): the volume of Reagent I (밀리리터) ~이다 1:5~10 to extract. It is recommended to add 1 mL of Reagent I to 0.1 g의 조직, and fully homogenize on ice bath. Centrifuge at 10000g for 10 4℃에서 1분 동안 불용성 물질 제거, 시험하기 전에 얼음에 상등액을 채취하십시오..

2. 박테리아 또는 세포

According to the bacteria or cells (104): the volume of Reagent I (밀리리터) 500~1000 입니다:1. It is recommended to add 1 mL of Reagent I to 5 수백만 개의 박테리아 또는 세포. 초음파 처리를 사용하여 박테리아와 세포를 분리합니다. (얼음 위에 놓음, ultrasonic power 300W, 근무 시간 3초, 간격 7초, 총 시간 3 분). 원심분리기, 10000g에 대한 10 4℃에서 1분간 가온하여 불용성 물질을 제거하고 상등액을 얼음에 취하여 시험한다.

3. Culture medium or other liquid: 직접 감지.

II. 발각

• 예열 분광 광도계 30 분, 파장을 조정하여 595 nm, 증류수로 0을 설정하다.
• Preheat reagent III at 30℃ for more than 20 분.
• 기준: Dilute the 10μmol/mL standard solution to 1.25, 0.625, 0.3125, 0.15625, 0.078μmol/mL with reagent I.
• 다음 시약을 추가하십시오 1.5 mL EP tubes:

	Contrast tube (씨)	시험관 (티)	표준 튜브 (에스)	빈 튜브 (비)
견본 (μL)	100	100	–	–
표준 용액 (μL)	–	–	100	–
시약 I (μL)	450	400	400	500
시약 II (μL)	–	50	50	50
Mix and react for 10 min at 30℃			–	–
시약 III (μL)	400	400	400	400
시약 IV (μL)	50	50	50	50
Mix thoroughly and detect the absorbance at 595 nm, record as AC, 에, AS and AB respectively. ΔAT=(AT-AC), ΔAS=(AS-AB). A contrast tube is required for each test tube, and the standard curve need only be tested once or twice.

III. 계산:

1.표준곡선

The concentration of standard solution as x-axis, ΔAS as y-axis, obtain the equation y=kx+b. Take ΔAT to the equation to acquire x (μmol/mL) value.

2. 계산

• 조직 단백질 농도

단위 정의: One unit of enzyme activity is defined as the amount of enzyme that catalyzes the hydrolysis of α-naphthyl acetate to generate 1 μmol of α-naphthol every mg of protein in the reaction system per minute at 40℃.

HL Activity (U/mg 프로트)=x×Vs÷（Vs×Cpr）÷T =0.1x÷ Cpr

• 조직 무게

단위 정의: One unit of enzyme activity is defined as the amount enzyme that catalyzes the hydrolysis of α-naphthyl acetate to generate 1 μmol of α-naphthol every gram of tissue in the reaction system per minute at 40℃.

HL Activity (U/g 중량) = x×Vs÷(W×Vs²Ve)÷T =0.0333x÷ W

• 액체

단위 정의: One unit of enzyme activity is defined as the amount of enzyme that catalyzes the hydrolysis of α-naphthyl acetate to generate 1 μmol of α-naphthol every milliliter of liquid sample in the reaction system per minute at 40℃.

HL Activity (U/mL) =x× Vs÷Vs÷T=0.1x

• 박테리아 또는 배양된 세포

단위 정의: One unit of enzyme activity is defined as the amount of enzyme that catalyzes the hydrolysis of α-naphthyl acetate to generate 1 μmol of α-naphthol every 104 cells or bacteria in the reaction system per minute at 40℃.

HL Activity (U/104 셀) =x× Ve÷ cell amount÷ T= 0.1x÷ cell amount

대: 샘플량 (밀리리터), 0.1 밀리리터; 베: 솔루션 볼륨 추출, 1 밀리리터;

심폐소생술: 상청액 샘플 단백질 농도 (mg/mL); 티: 반응 시간 (분), 10 분;

여: 샘플 중량, g;

셀량: 10 thousand as unit.

메모:

1. If the sample is animal liver, it is recommended to dilute the sample with reagent I more than 25 times before testing, and multiply the dilution factor in the calculation
2. If the sample is serum or plasma from obese animals, it is recommended to dilute the sample with reagent I more than 5 times before testing, and multiply the dilution factor in the calculation
3. When ΔA is greater than 0.8, it is recommended to measure the sample after diluting it with the reagent, and multiply it by the dilution factor in the calculation

실험예:

1. 1g rat liver was taken for sample processing, and the supernatant is diluted 24 타임스, then the operation is carried out according to the operation steps. Measured and calculated by 96 우물 접시: ΔA = AT-AB = 0.713-0.001=0.712, and the standard curve: y = 0.6381x – 0.0005, calculate x =1.1166

HL activity (U/g 질량) = x×VS ÷ (W×VS ÷ VST)÷ T ×48 = 53.597 U/g 질량.

2. After the turkey serum was diluted 6 타임스, the operation was carried out according to the operation steps. Measured and calculated by 96 우물 접시: ΔA = AT-AB =0.572-0.003=0.569, and the standard curve: y = 0.6381x – 0.0005, calculate x =892

HL activity (U/g 질량) = x×VS ÷ (W×VS ÷ VST)÷ T ×12 = 1.071 U/g 질량

관련 상품：

BC2340/BC2345 Lipase(LPS) 활동 분석 키트

BC0620/BC0625 Triglyceride(TG) 콘텐츠 분석 키트

BC2440/BC2445 Lipoprteinlipase(LPL) 활동 분석 키트

BC0320/BC0325 Plant Lipoxygenase(LOX) 활동 분석 키트

BC0590/BC0595 Free fatty Acids(FFA) 콘텐츠 분석 키트