Human Trappin-2/Elafin 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 Trappin-2/Elafin. 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: Trappin-2/Elafin
Trappin-2 is a member of the trappin family that includes the SLPI (secretory leukocyte protease inhibitor) protein. Elafin or ESI (elastase-specific inhibitor) and SKALP (skin-derived anti-leukoproteinase) are alternative names for trappin-2 and reflect its protease targets. However, Elafin and SKALP sometimes correspond only to the processed form that contains the C-terminal WAP domain of the molecule.
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 Trappin-2/Elafin 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.
16
Citations: Showing 1 - 10
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Effect of progestin-based contraceptives on HIV-associated vaginal immune biomarkers and microbiome in adolescent girls
Authors: Nasr, MA;Aldous, A;Daniels, J;Joy, C;Capozzi, E;Yang, M;Moriarty, P;Emmanuel-Baker, V;Malcolm, S;Green, SJ;Gomez-Lobo, V;Ghosh, M;
PloS one
Species: Human
Sample Types: Vaginal Swab
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Atypical response to bacterial co-infection and persistent neutrophilic broncho-alveolar inflammation distinguish critical COVID-19 from influenza
Authors: S Cambier, M Metzemaeke, AC Carvalho, A Nooyens, C Jacobs, L Vanderbeke, B Malengier-, M Gouwy, E Heylen, P Meersseman, G Hermans, E Wauters, A Wilmer, C Consortium, D Schols, P Matthys, G Opdenakker, RE Marques, J Wauters, J Vandooren, P Proost
JCI Insight, 2022-01-11;0(0):.
Species: Human
Sample Types: BAL Supernatant
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Addressing the challenges of E-cigarette safety profiling by assessment of pulmonary toxicological response in bronchial and alveolar mucosa models
Authors: K Ganguly, A Nordström, TA Thimraj, M Rahman, M Ramström, SI Sompa, EZ Lin, F O'Brien, J Koelmel, L Ernstgård, G Johanson, KJG Pollitt, L Palmberg, S Upadhyay
Sci Rep, 2020-11-24;10(1):20460.
Species: Human
Sample Types: Cell Culture Supernates
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Elafin inhibits obesity, hyperglycemia, and liver steatosis in high-fat diet-treated male mice
Authors: J Wang, C Ortiz, L Fontenot, R Mukhopadhy, Y Xie, IKM Law, DQ Shih, SA Mattai, Z Li, HW Koon
Sci Rep, 2020-07-30;10(1):12785.
Species: Human
Sample Types: Serum
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High circulating elafin levels are associated with Crohn's disease-associated intestinal strictures
Authors: J Wang, C Ortiz, L Fontenot, Y Xie, W Ho, SA Mattai, DQ Shih, HW Koon
PLoS ONE, 2020-04-14;15(4):e0231796.
Species: Human
Sample Types: Serum
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In-depth serum proteomics reveals biomarkers of psoriasis severity and response to traditional Chinese medicine
Authors: M Xu, J Deng, K Xu, T Zhu, L Han, Y Yan, D Yao, H Deng, D Wang, Y Sun, C Chang, X Zhang, J Dai, L Yue, Q Zhang, X Cai, Y Zhu, H Duan, Y Liu, D Li, Y Zhu, TRDJ Radstake, DMW Balak, D Xu, T Guo, C Lu, X Yu
Theranostics, 2019-04-13;9(9):2475-2488.
Species: Human
Sample Types: Serum
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A longitudinal analysis of the vaginal microbiota and vaginal immune mediators in women from sub-Saharan Africa
Authors: V Jespers, J Kyongo, S Joseph, L Hardy, P Cools, T Crucitti, M Mwaura, G Ndayisaba, S Delany-Mor, J Buyze, G Vanham, JHHM van de Wij
Sci Rep, 2017-09-20;7(1):11974.
Species: Human
Sample Types: Cervicovaginal Lavage Fluid
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Involvement of the CX3CL1 (fractalkine)/CX3CR1 pathway in the pathogenesis of acute graft-versus-host disease.
Authors: Brissot E, Bossard C, Malard F, Braudeau C, Chevallier P, Guillaume T, Delaunay J, Josien R, Gregoire M, Gaugler B, Mohty M
J Leukoc Biol, 2014-11-24;97(2):227-35.
Species: Human
Sample Types: Serum
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Aqueous two-phase system patterning of detection antibody solutions for cross-reaction-free multiplex ELISA.
Authors: Frampton, John P, White, Joshua B, Simon, Arlyne B, Tsuei, Michael, Paczesny, Sophie, Takayama, Shuichi
Sci Rep, 2014-05-02;4(0):4878.
Species: Human
Sample Types: Plasma
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Bone marrow mesenchymal stromal cells to treat tissue damage in allogeneic stem cell transplant recipients: correlation of biological markers with clinical responses.
Authors: Yin F, Battiwalla M, Ito S, Feng X, Chinian F, Melenhorst J, Koklanaris E, Sabatino M, Stroncek D, Samsel L, Klotz J, Hensel N, Robey P, Barrett A
Stem Cells, 2014-05-01;32(5):1278-88.
Species: Human
Sample Types: Plasma
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Searching for lower female genital tract soluble and cellular biomarkers: defining levels and predictors in a cohort of healthy Caucasian women.
Authors: Kyongo J, Jespers V, Goovaerts O, Michiels J, Menten J, Fichorova R, Crucitti T, Vanham G, Arien K
PLoS ONE, 2012-08-31;7(8):e43951.
Species: Human
Sample Types: Vaginal Lavage Fluid
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Selective impact of HIV disease progression on the innate immune system in the human female reproductive tract.
Authors: Lahey T, Ghosh M, Fahey JV, Shen Z, Mukura LR, Song Y, Cu-Uvin S, Mayer KH, Wright PF, Kappes JC, Ochsenbauer C, Wira CR
PLoS ONE, 2012-06-04;7(6):e38100.
Species: Human
Sample Types: Vaginal Lavage Fluid
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Trappin-2/elafin modulate innate immune responses of human endometrial epithelial cells to PolyI:C.
Authors: Drannik AG, Nag K, Yao XD, Henrick BM, Sallenave JM, Rosenthal KL
PLoS ONE, 2012-04-24;7(4):e35866.
Species: Human
Sample Types: Cell Culture Supernates
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Anti-HIV-1 activity of elafin is more potent than its precursor's, trappin-2, in genital epithelial cells.
Authors: Drannik AG, Nag K, Yao XD, Henrick BM, Jain S, Ball TB, Plummer FA, Wachihi C, Kimani J, Rosenthal KL
J. Virol., 2012-02-15;86(8):4599-610.
Species: Human
Sample Types: Vaginal Lavage Fluid
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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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Trappin-2/Elafin: a novel innate anti-human immunodeficiency virus-1 molecule of the human female reproductive tract.
Authors: Ghosh M, Shen Z, Fahey JV, Cu-Uvin S, Mayer K, Wira CR
Immunology, 2009-07-18;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Average Rating: 5 (Based on 2 Reviews)
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We used this kit for the quantification of Elafin in human serum and plasma. Works very well and well-described protocol.
