Human Periostin/OSF-2 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 Periostin/OSF-2. 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: Periostin/OSF-2
Periostin, also known as OSF-2, is a secreted matricellular protein with functions in extracellular matrix formation, cell migration, and inflammation. It is secreted as a 90 kDa monomer that can aggregate into >170 kDa higher-order multimers. Periostin is expressed by mesenchymal cells such as vascular smooth muscle cells, fibroblasts, osteoblasts, and odontoblasts in developing teeth. It is upregulated in many carcinomas. Periostin binds to multiple Integrins and enhances cell adhesion and cell migration. It enhances Fibronectin and Collagen I production and promotes collagen fibrillogenesis. It also induces epithelial-mesenchymal transition, tumor growth, invasion, and metastasis. Periostin induces the expression of VEGF R2 on endothelial cells and VEGF-C in tumor cells, and it can induce tumor lymphangiogenesis. Periostin plays an important role in heart valve development and tissue healing after myocardial infarction. In asthma, it is upregulated in bronchial epithelium and plays both destructive and protective roles by inducing eosinophil infiltration and inhibiting goblet cell metaplasia and mucus production.
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 Periostin/OSF-2 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.
31
Citations: Showing 1 - 10
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Effect of air pollutants on allergic inflammation in structural cells of nasal mucosa
Authors: Park, JH;Moon, JW;Yang, HW;Song, DJ;Park, IH;
Clinical and experimental otorhinolaryngology
Species: Human
Sample Types: Cell Culture Supernates
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Influence of diabetes mellitus on the pathological profile of aortic stenosis: a sex-based approach
Authors: Martín-Núñez, E;Goñi-Olóriz, M;Matilla, L;Garaikoetxea, M;Mourino-Alvarez, L;Navarro, A;Fernández-Celis, A;Tamayo, I;Gainza, A;Álvarez, V;Sádaba, R;Barderas, MG;Jover, E;López-Andrés, N;
Cardiovascular diabetology
Species: Human
Sample Types: Cell Culture Supernates, Tissue Homogenates
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Periostin deficiency attenuates kidney fibrosis in diabetic nephropathy by improving pancreatic ?-cell dysfunction and reducing kidney EMT
Authors: Cho, A;Jin, W;Lee, J;Shin, N;Lee, MS;Li, L;Yang, SH;Park, KS;Yang, CW;Kim, DK;Oh, YK;Lim, CS;Lee, JP;
Scientific reports
Species: Human
Sample Types: Urine
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Simple, low-cost, and well-performing method, the outgrowth technique, for the isolation of cells from nasal polyps
Authors: Kim, J;Hegener, K;Hagedorn, C;Weidinger, D;Jamal Jameel, K;Seuthe, IMC;Eichhorn, S;Kreppel, F;Park, JJ;Knobloch, J;
BMC molecular and cell biology
Species: Human
Sample Types: Cell Culture Supernates
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Abnormal basement membrane results in increased keratinocyte-derived periostin expression in psoriasis similar to wound healing
Authors: Flink, LB;Ghaffarinia, A;Papp, BT;Varga, Á;Vigh, AI;Vidács, DL;Kui, R;Kemény, L;Bata-Csörg?, Z;Bozó, R;
Scientific reports
Species: Human
Sample Types: Serum
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CTHRC1 Induces Pancreatic Stellate Cells (PSCs) into Myofibroblast-like Cancer-Associated Fibroblasts (myCAFs)
Authors: Kang, MK;Jiang, F;Kim, YJ;Ryu, K;Masamune, A;Hamada, S;Park, YY;Koh, SS;
Cancers
Species: Human
Sample Types: Cell Culture Supernates
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Long-Term Cryopreservation of Nasal Polyp Tissue in a Biobank for the Isolation and Culture of Primary Epithelial Cells
Authors: J Kim, K Hegener, C Hagedorn, K Jamal Jame, D Weidinger, IMC Seuthe, S Eichhorn, F Kreppel, J Knobloch, JJ Park
International Journal of Molecular Sciences, 2023-03-28;24(7):.
Species: Human
Sample Types: Cell Culture Supernates
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Nasal TSLP and periostin in infants with severe bronchiolitis and risk of asthma at 4�years of age
Authors: ML Garcia-Gar, B Sastre, M Arroyas, M Beato, P Alonso, JM Rodrigo-Mu, V Del Pozo, I Casas, C Calvo
Respiratory Research, 2023-01-24;24(1):26.
Species: Human
Sample Types: Nasopharyngeal aspirate
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Periostin Augments Vascular Smooth Muscle Cell Calcification via beta-Catenin Signaling
Authors: I Alesutan, LA Henze, B Boehme, TTD Luong, D Zickler, B Pieske, KU Eckardt, A Pasch, J Voelkl
Biomolecules, 2022-08-21;12(8):.
Species: Human
Sample Types: Serum
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Soluble POSTN is a novel biomarker complementing CA153 and CEA for breast cancer diagnosis and metastasis prediction
Authors: L Jia, G Li, N Ma, A Zhang, Y Zhou, L Ren, D Dong
BMC Cancer, 2022-07-12;22(1):760.
Species: Human
Sample Types: Cell Culture Supernates
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Serum Periostin Level and Genetic Polymorphisms Are Associated with Vertebral Fracture in Chinese Postmenopausal Women
Authors: YM Guo, JH Cheng, H Zhang, JW He, H Yue, WW Hu, JM Gu, YQ Hu, WZ Fu, C Wang, ZL Zhang
Genes, 2022-02-27;13(3):.
Species: Human
Sample Types: Serum
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Application of QPLEXTM biomarkers in cognitively normal individuals across a broad age range and diverse regions with cerebral amyloid deposition
Authors: D Lee, JC Park, KS Jung, J Kim, JS Jang, S Kwon, MS Byun, D Yi, G Byeon, G Jung, YK Kim, DY Lee, SH Han, I Mook-Jung
Experimental & Molecular Medicine, 2022-01-20;0(0):.
Species: Human
Sample Types: Plasma
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Involvement of the extracellular matrix proteins periostin and tenascin C in nasal polyp remodeling by regulating the expression of MMPs
Authors: K Du, M Wang, N Zhang, P Yu, P Wang, Y Li, X Wang, L Zhang, C Bachert
Clinical and translational allergy, 2021-09-06;11(7):e12059.
Species: Human
Sample Types: Tissue Homogenates
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Pathological Roles and Clinical Usefulness of Periostin in Type 2 Inflammation and Pulmonary Fibrosis
Authors: J Ono, M Takai, A Kamei, Y Azuma, K Izuhara
Biomolecules, 2021-07-22;11(8):.
Species: Human
Sample Types: Serum
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Expression profile of the matricellular protein periostin in paediatric inflammatory bowel disease
Authors: T Coelho, E Sonnenberg, Y Gao, E Mossotto, A Khojanazar, A Griffin, S Mukanova, A Ashimkhano, R Haggarty, A Borissenko, JJ Ashton, IS Stafford, A Batra, NA Afzal, MP Stanton, B Vadgama, K Adrisova, RM Beattie, AP Williams, S Ennis, D Riethmache
Scientific Reports, 2021-03-18;11(1):6194.
Species: Human
Sample Types: Plasma
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The potential pathways underlying the association of propyl-paraben exposure with aeroallergen sensitization and EASI score using metabolomics analysis
Authors: Y Lee, E Lee, DK Yon, HM Jee, HS Baek, SW Lee, JY Cho, MY Han
Scientific Reports, 2021-02-12;11(1):3772.
Species: Human
Sample Types: Serum
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iPSC-Derived Hereditary Breast Cancer Model Reveals the BRCA1-Deleted Tumor Niche as a New Culprit in Disease Progression
Authors: L Portier, C Desterke, D Chaker, N Oudrhiri, A Asgarova, F Dkhissi, AG Turhan, A Bennaceur-, F Griscelli
International Journal of Molecular Sciences, 2021-01-27;22(3):.
Species: Human
Sample Types: Cell Culture Supernates
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The vitamin D analogue calcipotriol promotes an anti-tumorigenic phenotype of human pancreatic CAFs but reduces T cell mediated immunity
Authors: L Gorchs, S Ahmed, C Mayer, A Knauf, C Fernández, M Svensson, R Heuchel, E Rangelova, P Bergman, H Kaipe
Sci Rep, 2020-10-15;10(1):17444.
Species: Human
Sample Types: Cell Culture Supernates
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Efficacy of endoscopic sinus surgery for eosinophilic chronic rhinosinusitis with asthma
Authors: N Ohta, Y Suzuki, H Ikeda, N Noguchi, R Kakuta, T Suzuki, R Ikeda, M Yamazaki, Y Saito, Y Kusano, Y Ishida, F Shoji, H Yoshioka, J Ono, T Takahashi, Y Nakamura, I Ohno, K Izuhara
Allergol Int, 2019-09-05;0(0):.
Species: Human
Sample Types: Serum
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A multiplex immunoassay of serum biomarkers for the detection of uveal melanoma
Authors: J Song, SL Merbs, LJ Sokoll, DW Chan, Z Zhang
Clin Proteomics, 2019-03-05;16(0):10.
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Bone Marrow Mesenchymal Stromal Cell-Derived Periostin Promotes B-ALL Progression by Modulating CCL2 in Leukemia Cells
Authors: Z Ma, X Zhao, M Deng, Z Huang, J Wang, Y Wu, D Cui, Y Liu, R Liu, G Ouyang
Cell Rep, 2019-02-05;26(6):1533-1543.e4.
Species: Human
Sample Types: Plasma
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Elevated serum periostin level in patients with chronic cough and airway hyperresponsiveness
Authors: Y Kono, M To, R Tsuzuki, K Haruki, Y To
Respir Investig, 2018-12-12;57(2):122-125.
Species: Human
Sample Types: Serum
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IL-13 induces periostin and eotaxin expression in human primary alveolar epithelial cells: Comparison with paired airway epithelial cells
Authors: Y Ito, R Al Mubarak, N Roberts, K Correll, W Janssen, J Finigan, R Mishra, HW Chu
PLoS ONE, 2018-04-19;13(4):e0196256.
Species: Human
Sample Types: Cell Culture Supernates
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Mixed Th2 and non-Th2 inflammatory pattern in the asthma-COPD overlap: a network approach
Authors: LP de Llano, BG Cosío, A Iglesias, N de Las Cue, JJ Soler-Cata, JL Izquierdo, JL López-Camp, C Calero, V Plaza, M Miravitlle, A Torrego, E Martinez-M, JB Soriano, AL Viña, I Bobolea
Int J Chron Obstruct Pulmon Dis, 2018-02-12;13(0):591-601.
Species: Human
Sample Types: Serum
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Asthma-COPD overlap is not a homogeneous disorder: further supporting data
Authors: L Pérez-de-L, BG Cosio
Respir. Res., 2017-11-02;18(1):183.
Species: Human
Sample Types: Serum
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Cigarette smoke differentially affects IL-13-induced gene expression in human airway epithelial cells
Authors: TCJ Mertens, AM van der Do, LE Kistemaker, DK Ninaber, C Taube, PS Hiemstra
Physiol Rep, 2017-07-11;5(13):.
Species: Human
Sample Types: Cell Culture Supernates
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Activated hepatic stellate cells secrete periostin to induce stem cell-like phenotype of residual hepatocellular carcinoma cells after heat treatment
Authors: R Zhang, RR Yao, JH Li, G Dong, M Ma, QD Zheng, DM Gao, JF Cui, ZG Ren, RX Chen
Sci Rep, 2017-05-19;7(1):2164.
Species: Human
Sample Types: Cell Culture Supernates
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Therapeutic Effect of Novel Single-Stranded RNAi Agent Targeting Periostin in Eyes with Retinal Neovascularization
Authors: T Nakama, S Yoshida, K Ishikawa, Y Kubo, Y Kobayashi, Y Zhou, S Nakao, T Hisatomi, Y Ikeda, K Takao, K Yoshikawa, A Matsuda, J Ono, S Ohta, K Izuhara, A Kudo, KH Sonoda, T Ishibashi
Mol Ther Nucleic Acids, 2017-02-09;6(0):279-289.
Species: Human
Sample Types: Cell Culture Supernates
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Circulating periostin in relation to insulin resistance and nonalcoholic fatty liver disease among overweight and obese subjects
Sci Rep, 2016-11-25;6(0):37886.
Species: Human
Sample Types: Serum
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Urinary Periostin Excretion Predicts Renal Outcome in IgA Nephropathy
Authors: Chun Soo Lim
Am. J. Nephrol., 2016-11-01;44(6):481-492.
Species: Human
Sample Types: Urine
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Polycyclic aromatic hydrocarbons reciprocally regulate IL-22 and IL-17 cytokines in peripheral blood mononuclear cells from both healthy and asthmatic subjects.
Authors: Ple C, Fan Y, Ait Yahia S, Vorng H, Everaere L, Chenivesse C, Balsamelli J, Azzaoui I, de Nadai P, Wallaert B, Lazennec G, Tsicopoulos A
PLoS ONE, 2015-04-10;10(4):e0122372.
Species: Human
Sample Types: Cell Culture Supernates
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sample dilution used 1:10