Human FABP3 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 Fatty Acid Binding Protein 3/FABP3. 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: FABP3/H-FABP
Fatty acid binding proteins are small cytoplasmic lipid binding proteins that are expressed in a tissue specific manner. FABPs bind free fatty acids, cholesterol, and retinoids, and are involved in intracellular lipid transport. Circulating FABP levels are used as indicators of tissue damage. Some FABP polymorphisms have been associated with disorders of lipid metabolism and the development of atherosclerosis.
Human FABP3, also known as FABP-Heart (H-FABP), FABP-Muscle (M-FABP), and mammary derived growth inhibitor (MDGI), is a member of the intracellular FABP family. FABP3 plays a role in the transport of long chain fatty acids in the cytoplasm.
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 FABP3 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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Dapagliflozin Improved Cardiac Function and Structure in Diabetic Patients with Preserved Ejection Fraction: Results of a Single Centre, Observational Prospective Study
Authors: Cortés, M;Lorenzo, O;Lumpuy-Castillo, J;Martínez-Albaladejo, S;Taibo-Urquía, M;Pello, AM;Bollas, AJ;Orejas, M;Navas, MÁ;Macia, E;Martínez, ME;Rueda, A;Tuñón, J;
Journal of clinical medicine
Species: Human
Sample Types: Plasma, Serum, Urine
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Loss of fatty acid binding protein 3 ameliorates lipopolysaccharide-induced inflammation and endothelial dysfunction
Authors: HC Nguyen, S Bu, S Nikfarjam, B Rasheed, DCR Michels, A Singh, S Singh, C Marszal, JJ McGuire, Q Feng, JC Frisbee, M Qadura, KK Singh
The Journal of Biological Chemistry, 2023-01-19;0(0):102921.
Species: Human
Sample Types: Cell Culture Supernates
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A Story of PA/BSA and Biomarkers to Diagnose Pulmonary Hypertension in Patients with Severe Aortic Valve Stenosis-The Rise of IGF-BP2 and GDF-15
Authors: J Kletzer, S Hecht, S Ramsauer, B Scharinger, R Kaufmann, J Kammler, J Kellermair, K Akbari, H Blessberge, C Steinwende, K Hergan, UC Hoppe, M Lichtenaue, E Boxhammer
Journal of cardiovascular development and disease, 2023-01-05;10(1):.
Species: Human
Sample Types: Plasma
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CT-Diagnosed Sarcopenia and Cardiovascular Biomarkers in Patients Undergoing Transcatheter Aortic Valve Replacement: Is It Possible to Predict Muscle Loss Based on Laboratory Tests?-A Multicentric Retrospective Analysis
Authors: S Hecht, E Boxhammer, R Kaufmann, B Scharinger, C Reiter, J Kammler, J Kellermair, M Hammerer, H Blessberge, C Steinwende, UC Hoppe, K Hergan, M Lichtenaue
Journal of personalized medicine, 2022-09-04;12(9):.
Species: Human
Sample Types: Plasma
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Peri-event plasma PCSK9 and hsCRP after an acute myocardial infarction correlate with early deterioration of left ventricular ejection fraction: a cohort study
Authors: LS Silva-Berm, A Vargas-Vil, CA Sánchez-Va, AC Palacio, AF Buitrago, CO Mendivil
Oncogene, 2022-07-21;21(1):61.
Species: Human
Sample Types: Plasma
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Expression of the Novel Cardiac Biomarkers sST2, GDF-15, suPAR, and H-FABP in HFpEF Patients Compared to ICM, DCM, and Controls
Authors: P Jirak, R Pistulli, M Lichtenaue, B Wernly, V Paar, LJ Motloch, R Rezar, C Jung, UC Hoppe, PC Schulze, D Kretzschma, RC Braun-Dull, T Bekfani
J Clin Med, 2020-04-15;9(4):.
Species: Human
Sample Types: Serum
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Novel Biomarkers in Patients with Chronic Kidney Disease: An Analysis of Patients Enrolled in the GCKD-Study
Authors: M Mirna, A Topf, B Wernly, R Rezar, V Paar, C Jung, H Salmhofer, K Kopp, UC Hoppe, PC Schulze, D Kretzschma, MP Schneider, UT Schultheis, C Sommerer, K Paul, G Wolf, M Lichtenaue, M Busch
J Clin Med, 2020-03-24;9(3):.
Species: Human
Sample Types: Plasma
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Propofol-based anaesthesia versus sevoflurane-based anaesthesia for living donor kidney transplantation: results of the VAPOR-1 randomized controlled trial
Authors: GJ Nieuwenhui, VB Nieuwenhui, MAJ Seelen, SP Berger, MC van den He, JGM Burgerhof, PJ Ottens, RJ Ploeg, HGD Leuvenink, MMRF Struys
Br J Anaesth, 2017-05-01;118(5):720-732.
Species: Human
Sample Types: Urine
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