Human Serum Albumin 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 Albumin. 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: Albumin
Albumin is the most abundant plasma protein, constituting approximately 60% of the total protein in blood. It is produced primarily by the liver and is important for the maintenance of colloidal osmotic blood pressure. Therefore, it maintains the balance of fluid between the vascular compartment and extravascular tissue. It also acts as a transport protein, carrying fatty acids, bilirubin, and various drugs and hormones.
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 Serum Albumin 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 - 10
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Intrathecal IgG and IgM synthesis correlates with neurodegeneration markers and corresponds to meningeal B cell presence in MS
Authors: Rodriguez-Mogeda, C;van Gool, MM;van der Mast, R;Nijland, R;Keasberry, Z;van de Bovekamp, L;van Delft, MA;Picon, C;Reynolds, R;Killestein, J;Teunissen, CE;de Vries, HE;van Egmond, M;Witte, ME;
Scientific reports
Species: Human
Sample Types: Serum, CSF
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Microplastic-induced hepatic adverse effects evaluated in advanced quadruple cell human primary models following three weeks of repeated exposure
Authors: Guraka, A;Souch, G;Duff, R;Brown, D;Moritz, W;Kermanizadeh, A;
Chemosphere
Species: Human
Sample Types: Cell Culture Supernates
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Effects of sevoflurane on lung epithelial permeability in experimental models of acute respiratory distress syndrome
Authors: Zhai, R;Lenga Ma Bonda, W;Leclaire, C;Saint-Béat, C;Theilliere, C;Belville, C;Coupet, R;Blondonnet, R;Bouvier, D;Blanchon, L;Sapin, V;Jabaudon, M;
Journal of translational medicine
Species: Mouse
Sample Types: BALF, Plasma
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Enhanced half-life and antitumor activity of Interleukin-15 through genetic fusion of a serum albumin-specific protein binder
Authors: D Kim, JH Park, TY Kim, DG Kim, JH Byun, HS Kim
International journal of pharmaceutics, 2022-07-26;625(0):122059.
Species: Transgenic Mouse
Sample Types: Serum
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Generation of proliferating human adult hepatocytes using optimized 3D culture conditions
Authors: S Rose, F Ezan, M Cuvellier, A Bruyère, V Legagneux, S Langouët, G Baffet
Scientific Reports, 2021-01-12;11(1):515.
Species: Human
Sample Types: Cell Culture Supernates
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Azithromycin ameliorates sulfur dioxide-induced airway epithelial damage and inflammatory responses
Authors: JP Joelsson, JA Kricker, AJ Arason, S Sigurdsson, B Valdimarsd, FR Gardarsson, CP Page, F Lehmann, T Gudjonsson, S Ingthorsso
Respir. Res., 2020-09-10;21(1):233.
Species: Mouse
Sample Types: BALF
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Resveratrol Reduced Liver Damage After Liver Resection in a Rat Model by Upregulating Sirtuin 1 (SIRT1) and Inhibiting the Acetylation of High Mobility Group Box 1 (HMGB1)
Authors: S Yu, X Zhou, H Xiang, S Wang, Z Cui, J Zhou
Med. Sci. Monit., 2019-05-01;25(0):3212-3220.
Species: Human
Sample Types: Cell Culture Supernates
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Selective Inactivation of Intracellular BiP/GRP78 Attenuates Endothelial Inflammation and Permeability in Acute Lung Injury
Authors: A Leonard, V Grose, AW Paton, JC Paton, DI Yule, A Rahman, F Fazal
Sci Rep, 2019-02-14;9(1):2096.
Species: Mouse
Sample Types: BALF
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Enhanced anticancer effect of oncostatin M combined with salinomycin in CD133+ HepG2 liver cancer cells
Authors: C Fu, L Wang, G Tian, C Zhang, Y Zhao, H Xu, M Su, Y Wang
Oncol Lett, 2018-12-05;17(2):1798-1806.
Species: Human
Sample Types: Cell Culture Supernates
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The VITamin D and OmegA-3 TriaL-Depression Endpoint Prevention (VITAL-DEP): Rationale and design of a large-scale ancillary study evaluating vitamin D and marine omega-3 fatty acid supplements for prevention of late-life depression
Authors: OI Okereke, CF Reynolds, D Mischoulon, G Chang, NR Cook, T Copeland, G Friedenber, JE Buring, JE Manson
Contemp Clin Trials, 2018-03-08;0(0):.
Species: Human
Sample Types: Serum
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Effect of hypotensive therapy combined with modified diet or zinc supplementation on biochemical parameters and mineral status in hypertensive patients
Authors: J Suliburska, K Skrypnik, M Szuli?ska, J Kupsz, P Bogda?ski
J Trace Elem Med Biol, 2018-02-18;47(0):140-148.
Species: Human
Sample Types: Serum
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RAGE inhibition reduces acute lung injury in mice
Authors: R Blondonnet, J Audard, C Belville, G Clairefond, J Lutz, D Bouvier, L Roszyk, C Gross, M Lavergne, M Fournet, L Blanchon, C Vachias, C Damon-Soub, V Sapin, JM Constantin, M Jabaudon
Sci Rep, 2017-08-03;7(1):7208.
Species: Human
Sample Types: Serum
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Secretion of the endoplasmic reticulum stress protein, GRP78, into the BALF is increased in cigarette smokers
Authors: MO Aksoy, V Kim, WD Cornwell, TJ Rogers, B Kosmider, K Bahmed, C Barrero, S Merali, N Shetty, SG Kelsen
Respir. Res., 2017-05-02;18(1):78.
Species: Human
Sample Types: BALF
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Induction of highly functional hepatocytes from human umbilical cord mesenchymal stem cells by HNF4alpha transduction.
Authors: Hang H, Yu Y, Wu N, Huang Q, Xia Q, Bian J
PLoS ONE, 2014-08-19;9(8):e104133.
Species: Human
Sample Types: Cell Culture Supernates
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