Angiotensin Converting Enzyme Kit

Instruction manual

First Edition (Revised on April, 2016)

[ INTENDED USE ]

The kit applies to measure angiotensin converting enzyme (ACE) activity in serum or plasma sample.

 

[ REAGENTS AND MATERIALS PROVIDED ]

Reagents	Quantity	Reagents	Quantity
Reagent I	3×50mL	Standard	1×100μL
Instruction manual	1

[ MATERIALS REQUIRED BUT NOT SUPPLIED ]

1. A biochemical analyzer or spectrophotometer capable of measuring absorbance at 340nm ± 10nm filter.

2. Glass cuvettes of 12.5px light path.

3. Water bath.

4. Transferpettor and tips.

5. Vortex mixer.

6. Test tube.

7. A source of pure water (preferably double distilled water and double distilled water).

 

[ STORAGE OF THE KITS ]

Reagent I: Buffer solution FAPGG. Preserved at 2-8°C for 6 months without light struck.

Standard: Angiotensin converting enzyme solution. Preserved at 2-8°C for 6 months without light struck.

 

[ SAMPLE REQUIRMENT ]

Separate serum and plasma as soon as possible after blood collection to avoid hemolysis. Plasma can

only be anticoagulated with heparin, not with EDTA.

 

[ ASSAY PROCEDURE ]

1. Automatic biochemical analyze

        I.Parameter setting

Main wavelength	340nm	Auxiliary wavelength	450nm
Reaction temperature	37°C	Sample	25μL
Reaction type	Rate method	Reaction direction	Down reaction (-)

II .Operation steps

Compositions	Blank	Sample
Reagent I	200μl	200μl
Water	25μl	-
Sample	-	25μl
Mix fully and incubate for 180 s at 37℃. Measure the OD value for 300 s at 340 nm after zero setting and then calculate the △A/min.

2. Spectrophotometry

    Operation steps

Compositions	Blank	Sample
Reagent I	1000μl	1000μl
Water	125μl	-
Sample	-	125μl
Mix fully and incubate for 180 s at 37°C. Measure the OD value for 300 s at 340 nm after zero setting and then calculate the △A/min.

[ FORMULA ]

1. Calibration with calibrator

 

2. Calculation with calculating factors:

Note：U/ml to U/L conversion coefficient:1000

Optical path:1 cm

Millimolar extinction coefficient of FAPGG at 340nm: 0.989

 

[ TEST PRINCIPLE ]

ACE catalyze the hydrolysis of phenylalanyl bisglycine peptide to produce phenylalanyl compounds and diglycine peptide. The maximum absorption peak of FAP was at 340 nm. The activity of ACE in the sample can be calculated indirectly by measuring the absorbance change rate at 340 nm.

 

 

[ ASSAY SIGNIFICANCE ]

The main function of angiotensin-converting enzyme (ACE) is to transform angiotensin into angiotensin, and Angiotensin-converting enzyme (ACE) converts the hormone angiotensin I to the active vasoconstrictor angiotensin II, which can elevate blood pressure. ACE activity increases in most sarcoids patients, especially in active stage of disease. Serum ACE mainly secreted from pulmonary capillary endothelial cells. ACE may change significantly in acute lung injury. Serum ACE level can be used as an important index to measure the degree of lung injury caused by various factors.

 

[ PERFORMANCE INDEX ]

1. The absorbance of blank reagent: A340nm ≥ 0.700 (optical path =1 cm).

2. The absorbance change rate: △A/min (optical path =1 cm)  ≤ 0.005 

3. Linear range: 0~150 U/L, r ²≥ 0.990.

4. Sensitivity: The minimum detectable dose of this kit is typically less than 5.1 U/L

5. Accuracy: Relative deviation ≤ 15%.

6. Precision: intra-CV≤ 6%, inter-CV≤ 10%.

7. Stability: The valid of kit is 6 months when stored at 2~8°C in the dark. It is stable for 1 month

when stored at 2~8°C in the dark after opening.

[ IMPORTANT NOTES ]

1. This product is for scientific research use only, not for clinical diagnosis.

2. The amount of reagent and sample can be increased and decreased proportionately according to the

volume of cuvette.

3. Wear gloves and work clothes during experiment, the reagents should be avoided to contact with

skin and clothing. Wash immediately with plenty of water if contact it carelessly.

4. Do not mix or substitute reagents from one kit lot to another.

5. Biochemical analyzer or spectrophotometer with 340 ± 10nm filter.

6. Bilirubin has a significant inhibitory effect on ACE. A significantly lower results may be gotten in serum from severe jaundice patients .