Human Chemerin 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 Chemerin. 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: Chemerin
Chemerin, also known as Tazarotene-induced Gene 2, (Tig-2), is a member of the Cystatin superfamily. Chemerin is synthesized as a 163 aa precursor that contains a hydrophobic 20 aa N-terminal sequence, an intervening 137 aa Cystatin fold-containing domain, and a six aa C-terminal prosegment. It is activated by proteases of the coagulation, fibrinolytic, and inflammatory cascades. Chemerin is a chemoattractant for cells bearing the Chem R23 receptor, including plasmacytoid dendritic cells and tissue macrophages.
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 Chemerin 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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Relationships between Circulating Biomarkers and Body Composition Parameters in Patients with Metabolic Syndrome: A Community-Based Study
Authors: Tarabeih, N;Kalinkovich, A;Ashkenazi, S;Cherny, SS;Shalata, A;Livshits, G;
International journal of molecular sciences
Species: Human
Sample Types: Plasma
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High Glucose Promotes Inflammation and Weakens Placental Defenses against E. coli and S. agalactiae Infection: Protective Role of Insulin and Metformin
Authors: R Jiménez-Es, D Vargas-Alc, P Flores-Esp, AC Helguera-R, O Villavicen, I Mancilla-H, C Irles, YD Torres-Ram, MY Valdespino, P Velázquez-, R Zamora-Esc, M Islas-Lópe, C Carranco-S, L Díaz, V Zaga-Clave, A Olmos-Orti
International Journal of Molecular Sciences, 2023-03-09;24(6):.
Species: Human
Sample Types: Placental Supernates
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Plasma Chemerin Is Induced in Critically Ill Patients with Gram-Positive Infections
Authors: Amend, P;Mester, P;Schmid, S;Müller, M;Buechler, C;Pavel, V;
Biomedicines
Species: Human
Sample Types: Plasma
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Single-cell and spatial analysis reveal interaction of FAP+ fibroblasts and SPP1+ macrophages in colorectal cancer
Authors: J Qi, H Sun, Y Zhang, Z Wang, Z Xun, Z Li, X Ding, R Bao, L Hong, W Jia, F Fang, H Liu, L Chen, J Zhong, D Zou, L Liu, L Han, F Ginhoux, Y Liu, Y Ye, B Su
Nature Communications, 2022-04-01;13(1):1742.
Species: Human
Sample Types: Plasma
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Chemerin Overexpression in the Liver Protects against Inflammation in Experimental Non-Alcoholic Steatohepatitis
Authors: R Pohl, S Feder, EM Haberl, L Rein-Fisch, TS Weiss, M Spirk, A Bruckmann, N McMullen, CJ Sinal, C Buechler
Biomedicines, 2022-01-07;10(1):.
Species: Human
Sample Types: Cell Culture Supernates
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First trimester secreted Frizzled-Related Protein 4 and other adipokine serum concentrations in women developing gestational diabetes mellitus
Authors: JHN Schuitemak, RHJ Beernink, A Franx, TIFH Cremers, MPH Koster
PLoS ONE, 2020-11-18;15(11):e0242423.
Species: Human
Sample Types: Serum
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Growth and differentiation factor 15 is a biomarker for low back pain-associated disability
Authors: N Tarabeih, A Shalata, S Trofimov, A Kalinkovic, G Livshits
Cytokine, 2019-02-15;117(0):8-14.
Species: Human
Sample Types: Plasma
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Expression of human chemerin induces insulin resistance in the skeletal muscle but does not affect weight, lipid levels, and atherosclerosis in LDL receptor knockout mice on high-fat diet.
Authors: Becker M, Rabe K, Lebherz C, Zugwurst J, Goke B, Parhofer KG, Lehrke M, Broedl UC
Diabetes, 2010-08-19;59(11):2898-903.
Species: Human
Sample Types: Serum
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Integrated proteomic analysis of human cancer cells and plasma from tumor bearing mice for ovarian cancer biomarker discovery.
Authors: Pitteri SJ, JeBailey L, Faca VM, Thorpe JD, Silva MA, Ireton RC, Horton MB, Wang H, Pruitt LC, Zhang Q, Cheng KH, Urban N, Hanash SM, Dinulescu DM
PLoS ONE, 2009-11-19;4(11):e7916.
Species: Human
Sample Types: Plasma
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Insulin and metformin regulate circulating and adipose tissue chemerin.
Authors: Tan BK, Chen J, Farhatullah S, Adya R, Kaur J, Heutling D, Lewandowski KC, O'Hare JP, Lehnert H, Randeva HS
Diabetes, 2009-06-05;58(9):1971-7.
Species: Human
Sample Types: Serum
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Chemerin expression marks early psoriatic skin lesions and correlates with plasmacytoid dendritic cell recruitment.
Authors: Albanesi C, Scarponi C, Pallotta S, Daniele R, Bosisio D, Madonna S, Fortugno P, Gonzalvo-Feo S, Franssen JD, Parmentier M, De Pita O, Girolomoni G, Sozzani S
J. Exp. Med., 2008-12-29;206(1):249-58.
Species: Human
Sample Types: Cell Culture Supernates
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Chemerin is a novel adipokine associated with obesity and metabolic syndrome.
Authors: Bozaoglu K, Bolton K, McMillan J, Zimmet P, Jowett J, Collier G, Walder K, Segal D
Endocrinology, 2007-07-19;148(10):4687-94.
Species: Human
Sample Types: Plasma
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