Human Galectin-3 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.
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
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 equivalent
Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).
Blocking Buffer: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).
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), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent
Scientific Data
Product Datasheets
Preparation and Storage
Background: Galectin-3
The galectins constitute a large family of carbohydrate-binding proteins that function in many systems both intracellularly and following secretion. Galectins contain either one or two carbohydrate recognition domains (CRR) which mediate recognition of N-acetyl-lactosamine-containing glycoproteins. Some galectins exist in multiple isoforms due to alternative splicing. Individual galectins differ in their tissue distribution and in their carbohydrate-binding specificities.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody (to the working concentration stated in the product datasheet ) 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 each well of the microplate as recommended in the product datasheet. Incubate at room temperature for a minimum of 1 hour.
Note: The recommended Reagent Diluent typically contains 1% BSA. Some DuoSet Development Kits require alternative blocking agents, or for plates to be blocked overnight with a higher percentage of BSA, please see the product datasheet for details.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
PRECAUTION
The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.
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 (as recommended in the product datasheet), 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 Galectin-3 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.
15
Citations: Showing 1 - 10
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Myocardial transcriptomic analysis of diabetic patients with aortic stenosis: key role for mitochondrial calcium signaling
Authors: Cherpaz, M;Meugnier, E;Seillier, G;Pozzi, M;Pierrard, R;Leboube, S;Farhat, F;Vola, M;Obadia, JF;Amaz, C;Chalabreysse, L;May, C;Chanon, S;Brun, C;Givre, L;Bidaux, G;Mewton, N;Derumeaux, G;Bergerot, C;Paillard, M;Thibault, H;
Cardiovascular diabetology
Species: Human
Sample Types: Serum
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Neutrophils and galectin-3 defend mice from lethal bacterial infection and humans from acute respiratory failure
Authors: Das, S;Kaminski, TW;Schlegel, BT;Bain, W;Hu, S;Patel, A;Kale, SL;Chen, K;Lee, JS;Mallampalli, RK;Kagan, VE;Rajasundaram, D;McVerry, BJ;Sundd, P;Kitsios, GD;Ray, A;Ray, P;
Nature communications
Species: Human
Sample Types: Endotracheal Tube Aspirates
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Predictors of HIV rebound differ by timing of antiretroviral therapy initiation
Authors: Li, JZ;Melberg, M;Kittilson, A;Abdel-Mohsen, M;Li, Y;Aga, E;Bosch, RJ;Wonderlich, ER;Kinslow, J;Giron, LB;Di Germanio, C;Pilkinton, M;MacLaren, L;Keefer, M;Fox, L;Barr, L;Acosta, E;Ananworanich, J;Coombs, R;Mellors, J;Deeks, S;Gandhi, RT;Busch, M;Landay, A;Macatangay, B;Smith, DM;AIDS Clinical Trials Group A5345 Study Team, ;
JCI insight
Species: Human
Sample Types: Plasma
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Vacuole Membrane Protein 1 (VMP1) Restricts NLRP3 Inflammasome Activation by Modulating SERCA Activity and Autophagy
Authors: SR Zack, R Nikolaienk, B Cook, R Melki, AV Zima, EM Campbell
Research square, 2023-01-27;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Biomarkers of Pulmonary Hypertension Are Altered in Children With Down Syndrome and Pulmonary Hypertension
Authors: M Griffiths, J Yang, D Vaidya, M Nies, S Brandal, DD Ivy, F Hickey, K Wolter-War, ED Austin, M Mullen, MW Pauciulo, KA Lutz, EB Rosenzweig, R Hirsch, D Yung, WC Nichols, AD Everett
The Journal of pediatrics, 2021-10-21;0(0):.
Species: Human
Sample Types: Plasma
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Beneficial Effects of Mineralocorticoid Receptor Pathway Blockade against Endothelial Inflammation Induced by SARS-CoV-2 Spike Protein
Authors: E Jover, L Matilla, M Garaikoetx, A Fernández-, P Muntendam, F Jaisser, P Rossignol, N López-Andr
Biomedicines, 2021-06-03;9(6):.
Species: Human
Sample Types: Cell Culture Supernates
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Exploring the potential effect of paricalcitol on markers of inflammation in de novo renal transplant recipients
Authors: HK Pihlstrøm, T Ueland, AE Michelsen, P Aukrust, F Gatti, C Hammarströ, M Kasprzycka, J Wang, G Haraldsen, G Mjøen, DO Dahle, K Midtvedt, IA Eide, A Hartmann, H Holdaas
PLoS ONE, 2020-12-16;15(12):e0243759.
Species: Human
Sample Types: Plasma
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Network analysis of the progranulin-deficient mouse brain proteome reveals pathogenic mechanisms shared in human frontotemporal dementia caused by GRN mutations
Authors: M Huang, E Modeste, E Dammer, P Merino, G Taylor, DM Duong, Q Deng, CJ Holler, M Gearing, D Dickson, NT Seyfried, T Kukar
Acta Neuropathol Commun, 2020-10-07;8(1):163.
Species: Human
Sample Types: Tissue Homogenates
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The Phenotype and Secretory Activity of Adipose-Derived Mesenchymal Stem Cells (ASCs) of Patients with Rheumatic Diseases
Authors: E Kuca-Warna, U Skalska, I Janicka, U Musia?owic, K Bonek, P G?uszko, P Szcz?sny, M Olesi?ska, E Kontny
Cells, 2019-12-17;8(12):.
Species: Human
Sample Types: Cell Culture Supernates
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Fecal Galectin-3: A New Promising Biomarker for Severity and Progression of Colorectal Carcinoma
Authors: M Jovanovic, N Gajovic, N Zdravkovic, M Jovanovic, M Jurisevic, D Vojvodic, V Maric, A Arsenijevi, I Jovanovic
Mediators Inflamm., 2018-04-04;2018(0):8031328.
Species: Human
Sample Types: Feces
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Galectin-3 acts as an angiogenic switch to induce tumor angiogenesis via Jagged-1/Notch activation
Authors: SN Dos Santos, H Sheldon, JX Pereira, C Paluch, EM Bridges, MC El-Cheikh, AL Harris, ES Bernardes
Oncotarget, 2017-07-25;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Increased expression of colonic Wnt9A through Sp1-mediated transcriptional effects involving arylsulfatase B, chondroitin 4-sulfate, and galectin-3.
Authors: Bhattacharyya S, Feferman L, Tobacman J
J Biol Chem, 2014-04-28;289(25):17564-75.
Species: Human
Sample Types: Cell Lysates
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Hypoxia reduces arylsulfatase B activity and silencing arylsulfatase B replicates and mediates the effects of hypoxia.
Authors: Bhattacharyya S, Tobacman JK
PLoS ONE, 2012-03-13;7(3):e33250.
Species: Human
Sample Types: Cell Lysates
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Human breast cancer cells enhance self tolerance by promoting evasion from NK cell antitumor immunity.
Authors: Mamessier E, Sylvain A, Thibult ML, Houvenaeghel G, Jacquemier J, Castellano R, Goncalves A, Andre P, Romagne F, Thibault G, Viens P, Birnbaum D, Bertucci F, Moretta A, Olive D
J. Clin. Invest., 2011-08-15;121(9):3609-22.
Species: Human
Sample Types: Cell Culture Supernates
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Novel INTeraction of MUC4 and galectin: potential pathobiological implications for metastasis in lethal pancreatic cancer.
Authors: Senapati S, Chaturvedi P, Chaney WG, Chakraborty S, Gnanapragassam VS, Sasson AR, Batra SK
Clin. Cancer Res., 2010-11-08;17(2):267-74.
Species: Human
Sample Types: Serum
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We used this kit to quantify Galectin 3 in human serum and plasma, works well.