Human S100B 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 S100 Calcium Binding Protein B (S100B). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma 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
Wash Buffer: (Catalog # WA126), or equivalent
Reagent Diluent*
Blocking Buffer*
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
*For the Reagent Diluent and Blocking Buffer recommended for a specific DuoSet ELISA Development Kit, please see the product.
Scientific Data
Product Datasheets
Preparation and Storage
Background: S100B
S100B belongs to the S100 subgroup of the EF-hand family of calcium binding proteins. It is a homodimer that is expressed primarily in the brain by astrocytes, oligodendrocytes and Schwann cells. S100B has multiple intracellular functions, but can also be secreted from cells to exert extracellular functions. Some of the extracellular functions of S100B may be mediated by RAGE (receptor for advanced glycation end products). Blood levels of S100B can be used to monitor the extent of brain injury and malignant melanoma.
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 Block Buffer 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 S100B 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.
14
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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Differential Expression of Circulating Damage-Associated Molecular Patterns in Patients with Coronary Artery Ectasia
Authors: Tsoporis, JN;Triantafyllis, AS;Kalogeropoulos, AS;Izhar, S;Rigopoulos, AG;Rallidis, LS;Sakadakis, E;Toumpoulis, IK;Salpeas, V;Leong-Poi, H;Parker, TG;Rizos, I;
Biomolecules
Species: Human
Sample Types: Plasma
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Cognitive dysfunction in systemic lupus erythematosus is associated with disease activity and oxidative stress: a comparative study with rheumatoid arthritis for identifying biomarkers
Authors: de Sousa, DC;Sobreira, EST;Feitosa, WLQ;Aires, TMPM;Araújo, LPP;Silva, ALC;Joventino, CB;Silveira, NMT;Chaves-Filho, AJM;Macêdo, DS;Braga-Neto, P;
BMC neuroscience
Species: Human
Sample Types: Plasma
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Serum Levels of S100B Protein and Myelin Basic Protein as a Potential Biomarkers of Recurrent Depressive Disorders
Authors: Levchuk, LA;Roschina, OV;Mikhalitskaya, EV;Epimakhova, EV;Simutkin, GG;Bokhan, NA;Ivanova, SA;
Journal of personalized medicine
Species: Human
Sample Types: Plasma
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Astrocytes mediate cell non-autonomous correction of aberrant firing in human FXS neurons
Authors: SD Sharma, BK Reddy, R Pal, TE Ritakari, JD Cooper, BT Selvaraj, PC Kind, S Chandran, DJA Wyllie, S Chattarji
Cell Reports, 2023-04-04;42(4):112344.
Species: Human
Sample Types: Cell Culture Supernates
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Effects of Exercise Training on Neurotrophic Factors and Blood-Brain Barrier Permeability in Young-Old and Old-Old Women
Authors: SY Cho, HT Roh
International Journal of Environmental Research and Public Health, 2022-12-16;19(24):.
Species: Human
Sample Types: Serum
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Carbon monoxide (CO) correlates with symptom severity, autoimmunity, and responses to probiotics treatment in a cohort of children with autism spectrum disorder (ASD): a post-hoc analysis of a randomized controlled trial
Authors: HT Sherman, K Liu, K Kwong, ST Chan, AC Li, XJ Kong
BMC Psychiatry, 2022-08-08;22(1):536.
Species: Human
Sample Types: Serum
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ProBDNF as an Indicator of Improvement among Women with Depressive Episodes
Authors: W Zwoli?ska, M Skibinska, A S?opie?, M Dmitrzak-W
Metabolites, 2022-04-16;12(4):.
Species: Human
Sample Types: Serum
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Longitudinal assessment of S100B serum levels and clinical factors in youth patients with mood disorders
Authors: A Rajewska-R, M Dmitrzak-W, P Kapelski, N Lepczynska, J Pawlak, J Twarowska-, M Skibinska
Scientific Reports, 2021-06-07;11(1):11973.
Species: Human
Sample Types: Serum
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Circulating Ligands of the Receptor for Advanced Glycation End Products and the Soluble Form of the Receptor Modulate Cardiovascular Cell Apoptosis in Diabetes
Authors: JN Tsoporis, E Hatziagela, S Gupta, S Izhar, V Salpeas, A Tsiavou, AG Rigopoulos, AS Triantafyl, JC Marshall, TG Parker, IK Rizos
Molecules, 2020-11-10;25(22):.
Species: Human
Sample Types: Plasma
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Severe Traumatic Brain Injury (TBI) Modulates the Kinetic Profile of the Inflammatory Response of Markers for Neuronal Damage
Authors: CR Schindler, T Lustenberg, M Woschek, P Störmann, D Henrich, P Radermache, I Marzi
J Clin Med, 2020-06-01;9(6):.
Species: Human
Sample Types: Serum
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Chronic Food Antigen-specific IgG-mediated Hypersensitivity Reaction as A Risk Factor for Adolescent Depressive Disorder
Authors: R Tao, Z Fu, L Xiao
Genomics Proteomics Bioinformatics, 2019-06-21;0(0):.
Species: Human
Sample Types: Serum
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Effects of Kudiezi Injection on Serum Inflammatory Biomarkers in Patients with Acute Cerebral Infarction
Authors: X Liu, X Jin, B Chen, X Liu, X Liang, X Fang, H Wu, X Fu, H Zheng, X Ding, N Duan, Y Zhang
Dis. Markers, 2018-09-02;2018(0):7936736.
Species: Human
Sample Types: Serum
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Cognitive dysfunction correlates with elevated serum S100B concentration in drug-free acutely relapsed patients with schizophrenia
Authors: Yunlong Tan
Psychiatry Res, 2016-09-22;247(0):6-11.
Species: Human
Sample Types: Serum
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Protocol was clear and standards worked, however we weren't able to get detectable levels in plasma
Samples applied neat, consistent replicates