Mouse 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 Mouse 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.
19
Citations: Showing 1 - 10
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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: Mouse
Sample Types: BALF
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Galectin-3 and Autophagy in Renal Acute Tubular Necrosis
Authors: Al-Salam, S;Jagadeesh, GS;Sudhadevi, M;Yasin, J;
International journal of molecular sciences
Species: Mouse
Sample Types: Tissue Homogenates
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Mutation of the galectin-3 glycan-binding domain (Lgals3-R200S) enhances cortical bone expansion in male mice and trabecular bone mass in female mice
Authors: KA Maupin, CR Diegel, PD Stevens, D Dick, VAI Vivari, BO Williams
FEBS Open Bio, 2022-09-14;0(0):.
Species: Transgenic Mouse
Sample Types: Plasma
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Galectin-3 inhibitor GB0139 protects against acute lung injury by inhibiting neutrophil recruitment and activation
Authors: DC Humphries, R Mills, C Boz, BJ McHugh, N Hirani, AG Rossi, A Pedersen, HT Schambye, RJ Slack, H Leffler, UJ Nilsson, W Wang, T Sethi, AC Mackinnon
Frontiers in Pharmacology, 2022-08-08;13(0):949264.
Species: Mouse
Sample Types: BALF
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Inhibition of Galectin-3 Impairs Antifungal Immune Response in Fungal Keratitis
Authors: Y Xiao, J Yang, Z Fu, Z Xiong, C Zhang, D He, Z Zhou, N Li, J Yuan
Disease Markers, 2022-04-09;2022(0):8316004.
Species: Mouse
Sample Types: Tissue Homogenates
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Gal3 Plays a Deleterious Role in a Mouse Model of Endotoxemia
Authors: JC Fernández-, AM Espinosa-O, I García-Dom, I Rosado-Sán, YM Pacheco, R Moyano, JG Monterde, JL Venero, RM de Pablos
International Journal of Molecular Sciences, 2022-01-21;23(3):.
Species: Mouse
Sample Types: Serum
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Oleanolic acid ameliorates intestinal alterations associated with EAE
Authors: B Gutierrez, I Gallardo, L Ruiz, Y Alvarez, V Cachofeiro, A Margolles, M Hernandez, ML Nieto
J Neuroinflammation, 2020-11-27;17(1):363.
Species: Mouse
Sample Types: Serum
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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: Mouse
Sample Types: Tissue Homogenates
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Galectin-3 enhances neutrophil motility and extravasation into the airways during Aspergillus fumigatus�infection
Authors: BD Snarr, G St-Pierre, B Ralph, M Lehoux, Y Sato, A Rancourt, T Takazono, SR Baistrocch, R Corsini, MP Cheng, M Sugrue, LR Baden, K Izumikawa, H Mukae, JR Wingard, IL King, M Divangahi, MS Satoh, BG Yipp, S Sato, DC Sheppard
PLoS Pathog., 2020-08-04;16(8):e1008741.
Species: Mouse
Sample Types: Serum
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Myocardial Ischemia Reperfusion Injury: Apoptotic, Inflammatory and Oxidative Stress Role of Galectin-3
Authors: S Al-Salam, S Hashmi
Cell. Physiol. Biochem., 2018-10-24;50(3):1123-1139.
Species: Mouse
Sample Types: Tissue Homogenates
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Osteopontin deficiency ameliorates Alport pathology by preventing tubular metabolic deficits
Authors: W Ding, K Yousefi, S Goncalves, BJ Goldstein, AL Sabater, A Kloosterbo, P Ritter, G Lambert, AJ Mendez, LA Shehadeh
JCI Insight, 2018-03-22;3(6):.
Species: Mouse
Sample Types: Plasma
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Galectin-3 Plays an Important Role in Preterm Birth Caused by Dental Infection of Porphyromonas gingivalis
Authors: M Miyauchi, M Ao, H Furusho, C Chea, A Nagasaki, S Sakamoto, T Ando, T Inubushi, K Kozai, T Takata
Sci Rep, 2018-02-12;8(1):2867.
Species: Mouse
Sample Types: Amniotic Fluid
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Tsc2 disruption in mesenchymal progenitors results in tumors with vascular anomalies overexpressing Lgals3
Authors: PJ Klover, RL Thangapazh, J Kato, JA Wang, SA Anderson, V Hoffmann, WK Steagall, S Li, E McCart, N Nathan, JD Bernstock, MD Wilkerson, CL Dalgard, J Moss, TN Darling
Elife, 2017-07-11;6(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Effects of carvedilol on structural and functional outcomes and plasma biomarkers in the mouse transverse aortic constriction heart failure model
Authors: C Hampton, R Rosa, D Szeto, G Forrest, B Campbell, R Kennan, S Wang, CH Huang, L Gichuru, X Ping, X Shen, K Small, J Madwed, JJ Lynch
SAGE Open Med, 2017-03-30;5(0):2050312117700.
Species: Mouse
Sample Types: Plasma
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Aldosterone induced galectin-3 secretion in vitro and in vivo: from cells to humans.
Authors: Lin Y, Chou C, Wu X, Chang Y, Hung C, Chen Y, Tzeng Y, Wu V, Ho Y, Hsieh F, Wu K
PLoS ONE, 2014-09-02;9(9):e95254.
Species: Mouse
Sample Types: Cell Culture Supernates
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Free fatty acids and IL-6 induce adipocyte galectin-3 which is increased in white and brown adipose tissues of obese mice.
Authors: Krautbauer S, Eisinger K, Hader Y, Buechler C
Cytokine, 2014-07-17;69(2):263-71.
Species: Mouse
Sample Types: Cell Culture Supernates
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Obesity and IL-6 interact in modulating the response to endotoxemia in mice.
Cytokine, 2012-09-23;61(1):71-7.
Species: Mouse
Sample Types: Plasma
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Rosiglitazone improves survival and hastens recovery from pancreatic inflammation in obese mice.
Authors: Pini M, Rhodes DH, Castellanos KJ
PLoS ONE, 2012-07-16;7(7):e40944.
Species: Mouse
Sample Types: Plasma
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Galectin-3 is a negative regulator of lipopolysaccharide-mediated inflammation.
Authors: Li Y, Komai-Koma M, Gilchrist DS, Hsu DK, Liu FT, Springall T, Xu D
J. Immunol., 2008-08-15;181(4):2781-9.
Species: Mouse
Sample Types: Serum
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