Human ACE DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human ACE. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Scientific Data
Product Datasheets
Preparation and Storage
Background: ACE/CD143
ACE and ACE-2, two cell surface proteases, are important regulators of the renin-angiotensin system (RAS), which plays a key role in maintaining blood pressure homeostasis and fluid salt balance in mammals. Because of its location and specificity, ACE plays additional roles in immunity, reproduction and neuropeptide regulation. ACE exists in two isoforms. Somatic ACE comprises two highly similar protease domains whereas germinal ACE comprises a single protease domain. ACE-2 consists of a single protease domain and is an essential regulator of heart and lung function. It is also a cellular receptor for the Spike protein of SARS cornonavirus that causes severe acute respiratory syndrome.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Human ACE DuoSet ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 8
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Application of QPLEXTM biomarkers in cognitively normal individuals across a broad age range and diverse regions with cerebral amyloid deposition
Authors: D Lee, JC Park, KS Jung, J Kim, JS Jang, S Kwon, MS Byun, D Yi, G Byeon, G Jung, YK Kim, DY Lee, SH Han, I Mook-Jung
Experimental & Molecular Medicine, 2022-01-20;0(0):.
Species: Human
Sample Types: Plasma
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Human Tissue Angiotensin Converting Enzyme (ACE) Activity Is Regulated by Genetic Polymorphisms, Posttranslational Modifications, Endogenous Inhibitors and Secretion in the Serum, Lungs and Heart
Authors: V Bánhegyi, A Enyedi, GÁ Fülöp, A Oláh, IM Siket, C Váradi, K Bottyán, M Lódi, A Csongrádi, AJ Umar, M Fagyas, D Czuriga, I Édes, M Pólos, B Merkely, Z Csanádi, Z Papp, G Szabó, T Radovits, I Takács, A Tóth
Cells, 2021-07-06;10(7):.
Species: Human
Sample Types: Serum
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Angiotensin Converting Enzyme (ACE): A Marker for Personalized Feedback on Dieting
Authors: S Tejpal, N Sanghera, V Manoharan, J Planas-Igl, CC Bastie, J Klein-Seet
Nutrients, 2020-02-28;12(3):.
Species: Human
Sample Types: Urine
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Benefits of whole body vibration training in patients hospitalised for COPD exacerbations - a randomized clinical trial.
Authors: Greulich T, Nell C, Koepke J, Fechtel J, Franke M, Schmeck B, Haid D, Apelt S, Filipovic S, Kenn K, Janciauskiene S, Vogelmeier C, Koczulla A
BMC Pulm Med, 2014-04-11;14(0):60.
Species: Human
Sample Types: Serum
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New perspectives in the renin-angiotensin-aldosterone system (RAAS) IV: circulating ACE2 as a biomarker of systolic dysfunction in human hypertension and heart failure.
Authors: Uri K, Fagyas M, Manyine Siket I, Kertesz A, Csanadi Z, Sandorfi G, Clemens M, Fedor R, Papp Z, Edes I, Toth A, Lizanecz E
PLoS ONE, 2014-04-01;9(4):e87845.
Species: Human
Sample Types: Serum
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New perspectives in the renin-angiotensin-aldosterone system (RAAS) III: endogenous inhibition of angiotensin converting enzyme (ACE) provides protection against cardiovascular diseases.
Authors: Fagyas M, Uri K, Siket I, Darago A, Boczan J, Banyai E, Edes I, Papp Z, Toth A
PLoS ONE, 2014-04-01;9(4):e93719.
Species: Human
Sample Types: Serum
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Absence of cell surface expression of human ACE leads to perinatal death.
Authors: Michaud A, Acharya K, Masuyer G, Quenech'du N, Gribouval O, Moriniere V, Gubler M, Corvol P
Hum Mol Genet, 2013-10-24;23(6):1479-91.
Species: Human
Sample Types: Cell Lysates
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Changes with age in the activities of beta-secretase and the Abeta-degrading enzymes neprilysin, insulin-degrading enzyme and angiotensin-converting enzyme.
Authors: Miners JS, van Helmond Z, Kehoe PG, Love S
Brain Pathol., 2010-01-12;20(4):794-802.
Species: Human
Sample Types: Tissue Homogenates
FAQs
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Is ACE-1 the same as ACE?
Yes. ACE-1 is used as an alternative name, but not as commonly as ACE and CD143.
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