Human Pentraxin 3/TSG-14 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 Pentraxin 3. 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
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: Pentraxin 3/TSG-14
Pentraxin 3 (PTX3), also known as TNF-stimulated gene 14 (TSG-14), is a secreted glycoprotein belonging to the Pentraxin family of proteins. It is the prototypical long Pentraxin, exhibiting a C-terminal Pentraxin domain characteristic of the family, and a unique N-terminal domain. It is produced by several cell types including endothelial and mononuclear cells. PTX3 acts as a pattern recognition receptor with non-redundant roles in the innate immune response to several microbial agents including the fungal pathogen Aspergillus fumigatus. Overexpression leads to enhanced pro-inflammatory responses. PTX3 is induced in response to LPS and inflammatory cytokines including TNF-alpha and IL-1 beta.
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 Pentraxin 3/TSG-14 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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RNA m6A methylation modulates airway inflammation in allergic asthma via PTX3-dependent macrophage homeostasis
Authors: Han, X;Liu, L;Huang, S;Xiao, W;Gao, Y;Zhou, W;Zhang, C;Zheng, H;Yang, L;Xie, X;Liang, Q;Tu, Z;Yu, H;Fu, J;Wang, L;Zhang, X;Qian, L;Zhou, Y;
Nature communications
Species: Human
Sample Types: Serum, Cell Culture Supernates, Plasma
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Pentraxin 3 and Shigella LPS and IpaB Antibodies Interplay to Defeat Shigellosis
Authors: S Meron-Suda, A Reizis, S Goren, A Bialik, A Hochberg, D Cohen
Journal of Clinical Medicine, 2022-07-28;11(15):.
Species: Human
Sample Types: Serum
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Prediction of emergency cerclage outcomes in women with cervical insufficiency: The role of inflammatory, angiogenic, and extracellular matrix-related proteins in amniotic fluid
Authors: KN Lee, KH Park, YM Kim, I Cho, TE Kim
PLoS ONE, 2022-05-10;17(5):e0268291.
Species: Human
Sample Types: Amniotic Fluid
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Oxidative Stress-Induced Pentraxin 3 Expression Human Retinal Pigment Epithelial Cells is Involved in the Pathogenesis of Age-Related Macular Degeneration
Authors: N Hwang, MY Kwon, JM Woo, SW Chung
Int J Mol Sci, 2019-11-29;20(23):.
Species: Human
Sample Types: Cell Culture Supernates
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Loss of skin elasticity is associated with pulmonary emphysema, biomarkers of inflammation, and matrix metalloproteinase activity in smokers
Authors: ME O'Brien, D Chandra, RC Wilson, CM Karoleski, CR Fuhrman, JK Leader, J Pu, Y Zhang, A Morris, S Nouraie, J Bon, Z Urban, FC Sciurba
Respir. Res., 2019-06-24;20(1):128.
Species: Human
Sample Types: Plasma
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Calcineurin inhibitors reduce NFAT-dependent expression of antifungal pentraxin-3 by human monocytes
Authors: K Bendí?ková, F Tidu, M De Zuani, MH Kohoutková, I Andrej?ino, A Pompeiano, S B?lášková, G Forte, T Zelante, J Fri?
J. Leukoc. Biol., 2019-04-01;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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The Long Pentraxin 3 Contributes to Joint Inflammation in Gout by Facilitating the Phagocytosis of Monosodium Urate Crystals
Authors: NV Batista, M Barbagallo, VLS Oliveira, T Castro-Gom, RDR Oliveira, P Louzada-Ju, GRC Pinheiro, A Mantovani, MM Teixeira, C Garlanda, FA Amaral
J. Immunol., 2019-02-04;0(0):.
Species: Human
Sample Types: Plasma
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Role of a fluid-phase PRR in fighting an intracellular pathogen: PTX3 in Shigella infection
Authors: V Ciancarell, L Lembo-Fazi, I Paciello, AK Bruno, S Jaillon, S Berardi, M Barbagallo, S Meron-Suda, D Cohen, A Molinaro, G Rossi, C Garlanda, ML Bernardini
PLoS Pathog., 2018-12-07;14(12):e1007469.
Species: Human
Sample Types: Cell Culture Supernates
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Serum Pentraxin 3 is associated with signs of arterial alteration in women with preeclampsia
Authors: T Akhter, AK Wikström, M Larsson, A Larsson, G Wikström, T Naessen
Int. J. Cardiol., 2017-03-24;0(0):.
Species: Human
Sample Types: Serum
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Tunicamycin-induced Endoplasmic Reticulum Stress Upregulates the Expression of Pentraxin 3 in Human Retinal Pigment Epithelial Cells
Authors: Narae Hwang
Korean J Ophthalmol, 2016-12-06;30(6):468-478.
Species: Human
Sample Types: Cell Culture Supernates
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Role of pentraxin 3 in shaping arthritogenic alphaviral disease: from enhanced viral replication to immunomodulation.
Authors: Foo, Suan-Sin, Chen, Weiqiang, Taylor, Adam, Sheng, Kuo-Chin, Yu, Xing, Teng, Terk-Shi, Reading, Patrick, Blanchard, Helen, Garlanda, Cecilia, Mantovani, Alberto, Ng, Lisa F P, Herrero, Lara J, Mahalingam, Suresh
PLoS Pathog, 2015-02-19;11(2):e1004649.
Species: Human
Sample Types: Serum
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Severe Acinetobacter baumannii sepsis is associated with elevation of pentraxin 3.
Authors: Ketter P, Guentzel M, Schaffer B, Herzig M, Wu X, Montgomery R, Parida B, Fedyk C, Yu J, Jorgensen J, Chambers J, Cap A, Arulanandam B
Infect Immun, 2014-07-07;82(9):3910-8.
Species: Mouse
Sample Types: Serum
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Inflammatory biomarker pentraxin 3 (PTX3) in relation to obesity, body fat depots and weight loss.
Authors: Witasp A, Carrero J, Michaelsson K, Ahlstrom H, Kullberg J, Adamsson V, Riserus U, Larsson A, Helmersson-Karlqvist J, Lind L, Stenvinkel P, Arnlov J
Obesity (Silver Spring), 2014-01-25;22(5):1373-9.
Species: Human
Sample Types: Serum
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Exogenous pentraxin 3 restores antifungal resistance and restrains inflammation in murine chronic granulomatous disease.
Authors: D'Angelo C, D'Angelo C, De Luca A, Zelante T, Bonifazi P, Moretti S, Giovannini G, Iannitti RG, Zagarella S, Bozza S, Campo S, Salvatori G, Romani L
J. Immunol., 2009-09-04;183(7):4609-18.
Species: Human
Sample Types: Tissue Homogenates
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