Angiotensin I Converting Enzyme 1 ELISA Kit

Produktnummer ABIN956177

Dieses Colorimetric ELISA-Kit wurde entwickelt für die quantitative Messung von Human Angiotensin I Converting Enzyme 1.

Kurzübersicht für Angiotensin I Converting Enzyme 1 ELISA Kit (ABIN956177)

Target: Angiotensin I Converting Enzyme 1 (ACE) (Angiotensin I Converting Enzyme (Peptidyl-Dipeptidase A) 1 (ACE))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Competition ELISA

Applikation: ELISA

Produktdetails Angiotensin I Converting Enzyme 1 ELISA Kit

Verwendungszweck: This ACE ELISA kit is intended for laboratory research use only and is not for use in diagnostic or therapeutic procedures.

Analytische Methode: Quantitative

Produktmerkmale: The coated well immunoenzymatic assay for the quantitative measurement of serum ACE utilizes a monoclonal anti-ACE and a ACE-HRP conjugate. The assay sample and buffer are incubated together with anti-ACE antibody coated plate for sixty and washed. The diluted ACE-HRP conjugate is then added to each well and incubated. After the incubation period, the wells are decanted and washed three times. The wells are then incubated with a substrate for the enzyme. The product of the enzyme-substrate reaction forms a blue colored complex. Finally, a stopping solution is added to stop the reaction, which will then turn the solution yellow. The intensity of color is measured spectrophotometrically at 450 nm in a microplate reader. The intensity of the color is inversely proportional to the ACE concentration since ACE from samples and ACE-HRP conjugate compete for the anti-ACE antibody binding site. Since the number of sites is limited, as more sites are occupied by ACE from the sample, fewer sites are left to bind ACE- HRP conjugate. Calibrators of known ACE concentrations are run concurrently with the samples being assayed and a calibration curve is plotted relating the intensity of the color (Optical Density) to the concentration of ACE. The unknown ACE concentration in each sample is interpolated from this curve.

Bestandteile: Microtiter Plate: 96 wells
Calibrator 1 (0 ng/mL)
Calibrator 2 (1 ng/mL)
Calibrator 3 (2.5 ng/mL)
Calibrator 4 (5 ng/mL)
Calibrator 5 (10 ng/mL)
Calibrator 6 (25 ng/mL)
Enzyme Conjugate (1 x 6 mL)
Substrate A (1 x 6 mL)
Substrate B 1 x 6 mL
Stop Solution 1 x 6 mL
Wash Buffer (100X concentrate) 1 x 10 mL
Lysis Buffer Solution 1 x 6 mL
Note: The lysis buffer solution is used only when the sample is cell culture fluid & body fluid & tissue homogenate, If the sample is serum or blood plasma, then the lysis buffer solution is a superfluous reagent.

Benötigtes Material: 1. Microplate reader capable of measuring absorbance at 450 nm.
2. Precision pipettes to deliver 2 mL to 1 mL volumes.
3. Adjustable 10-100 mL pipettes for reagent preparation.
4. 100 mL and 1 liter graduated cylinders.
5. Calibrated adjustable precision pipettes, preferably with disposable plastic tips. (A manifold multi- channel pipette is desirable for large assays.)
6. 37°C incubator.
7. Absorbent paper.
8. Distilled or de-ionized water
9. Data analysis and graphing software. Graph paper: linear (Cartesian), log-log or semi-log, or log-logit as desired.
10. Tubes to prepare calibrator or sample dilutions.

Anwendungsinformationen

Plattentyp: Pre-coated

Testdurchführung:

It is recommended that all Calibrators and Samples be added in duplicate to the Microtiter Plate.
1. Secure the desired number of coated wells in the holder, then add 100 µL of Calibrators or Samples to the appropriate well of the antibody pre-coated Microtiter Plate.
2. Add 50 µL of Conjugate to each well. Mix well. Complete mixing in this step is important. Cover and incubate for 1 hour at 37°C.
3. Wash the microtiter plate using one of the specified methods indicated below:
4. Manual Washing: Remove incubation mixture by aspirating contents of the plate into a sink or proper waste container. Using a squirt bottle, fill each well completely with wash solution, then aspirate contents of the plate into a sink or proper waste container. Repeat this procedure four more times for a total of five washes. After final wash, invert plate, and blot dry by hitting plate onto absorbent paper or paper towels until no moisture appears. Note: Hold the sides of the plate frame firmly when washing the plate to assure that all strips remain securely in frame.
5. Automated Washing: Aspirate all wells, then wash plate five times using wash solution. Always adjust your washer to aspirate as much liquid as possible and set fill volume at 350 µL/well/wash (range: 350-400 µL). After final wash, invert plate, and blot dry by hitting plate onto absorbent paper or paper towels until no moisture appears. It is recommended that the washer be set for a soaking time of 10 seconds or shaking time of 5 seconds between washes.
6. Add 50 µL substrate A and 50 µL substrate B to each well. Cover and incubate for 10 minutes at 20- 25°C. (Avoid exposure to light.)
7. Add 50 µL stop solution to each well. Mix well.
8. Read the optical density (OD) at 450 nm using a microtiter plate reader immediately.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: 4 °C

Informationen zur Lagerung: All reagents provided are stored at 4°C. Refer to the expiration date on the label.

Haltbarkeit: The expiry date is stated on the label.

Details zu Angiotensin I Converting Enzyme 1

Target: Angiotensin I Converting Enzyme 1 (ACE) (Angiotensin I Converting Enzyme (Peptidyl-Dipeptidase A) 1 (ACE))

Andere Bezeichnung: Angiotensin I Converting Enzyme (ACE)

Pathways: ACE Inhibitor Pathway, Peptide Hormone Metabolism, Regulation of Systemic Arterial Blood Pressure by Hormones, Feeding Behaviour, Smooth Muscle Cell Migration