Human CXCL7/NAP-2 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY393
Ancillary Products Available
Human CXCL7 / NAP-2 ELISA Standard Curve
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Product Details
Procedure
Citations (13)
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Human CXCL7/NAP-2 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
15.6 - 1,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* 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 CXCL7/NAP-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

Human CXCL7 / NAP-2 ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CXCL7/NAP-2

Neutrophil activating peptide 2 (NAP-2), connective tissue activating protein III (CTAP-III) and beta-thrombogulin (beta-TG), are proteolytically processed carboxy-terminal fragments of platelet basic protein (PBP), which is found in the alpha-granules of human platelets. NAP-2 is a member of the CXC chemokines and is named CXCL7. Similar to other ELR domain containing CXC chemokines, such as IL-8 and the GRO proteins, NAP-2 binds CXCR2 and chemoattracts and activates neutrophils.

Entrez Gene IDs:
5473 (Human)
Alternate Names:
Beta-TG; connective tissue-activating peptide III; CTAP III; CTAP3CTAP-III; CTAPIII; CXC chemokine ligand 7; C-X-C motif chemokine 7; CXCL7; CXCL7b-TG1; LA-PF4; LDGFTC1; Leukocyte-derived growth factor; low-affinity platelet factor IV; Macrophage-derived growth factor; MDGFTC2; NAP2; NAP-2; NAP-2-L1; neutrophil-activating peptide 2; neutrophil-activating peptide-2; PBPTHBGB; platelet basic protein; PPBP; pro-platelet basic protein (chemokine (C-X-C motif) ligand 7); SCAR10; SCYB7beta-thromboglobulin; small inducible cytokine B7; small inducible cytokine subfamily B, member 7; Small-inducible cytokine B7; TGB; TGB1B-TG1; THBGB1; thrombocidin 1; thrombocidin 2; thromboglobulin, beta-1

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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 CXCL7/NAP-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.

13 Citations: Showing 1 - 10
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  1. Development of an Electrochemical CCL5 Chemokine Immunoplatform for Rapid Diagnosis of Multiple Sclerosis
    Authors: S Guerrero, E Sánchez-Ti, L Agüí, A González-C, P Yáñez-Sede, JM Pingarrón
    Biosensors, 2022;12(8):.
    Species: Human
    Sample Types: Serum
  2. The effect of transdermal gender-affirming hormone therapy on markers of inflammation and hemostasis
    Authors: MH Schutte, R Kleemann, NM Nota, CM Wiepjes, JM Snabel, G T'Sjoen, A Thijs, M den Heijer
    PLoS ONE, 2022;17(3):e0261312.
    Species: Human
    Sample Types: Serum
  3. ADP secreted by dying melanoma cells mediates chemotaxis and chemokine secretion of macrophages via the purinergic receptor P2Y12
    Authors: L Kloss, C Dollt, K Schledzews, A Krewer, S Melchers, C Manta, C Sticht, C Torre, J Utikal, V Umansky, A Schmieder
    Cell Death Dis, 2019;10(10):760.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Inhibition of profibrotic microRNA-21 affects platelets and their releasate
    Authors: T Barwari, S Eminaga, U Mayr, R Lu, PC Armstrong, MV Chan, M Sahraei, M Fernández-, T Moreau, J Barallobre, M Lynch, X Yin, C Schulte, F Baig, R Pechlaner, SR Langley, A Zampetaki, P Santer, M Weger, R Plasenzott, M Schosserer, J Grillari, S Kiechl, J Willeit, AM Shah, C Ghevaert, TD Warner, C Fernández-, Y Suárez, M Mayr
    JCI Insight, 2018;3(21):.
    Species: Human
    Sample Types: Plasma
  5. A combination of platelet features allows detection of early-stage cancer
    Authors: S Sabrkhany, MJE Kuijpers, SMJ van Kuijk, L Sanders, S Pineda, SWM Olde Damin, AC Dingemans, AW Griffioen, MGA Oude Egbri
    Eur. J. Cancer, 2017;80(0):5-13.
    Species: Human
    Sample Types: Platelets
  6. Proteome-wide analysis and CXCL4 as a biomarker in systemic sclerosis
    Authors: L van Bon, AJ Affandi, J Broen, RB Christmann, RJ Marijnisse, L Stawski, GA Farina, G Stifano, AL Mathes, M Cossu, M York, C Collins, M Wenink, R Huijbens, R Hesselstra, T Saxne, M DiMarzio, D Wuttge, SK Agarwal, JD Reveille, S Assassi, M Mayes, Y Deng, JP Drenth, J de Graaf, M den Heijer, CG Kallenberg, M Bijl, A Loof, WB van den Be, LA Joosten, V Smith, F de Keyser, R Scorza, C Lunardi, PL van Riel, M Vonk, W van Heerde, S Meller, B Homey, L Beretta, M Roest, M Trojanowsk, R Lafyatis, TR Radstake
    N. Engl. J. Med, 2014;370(5):433-43.
    Species: Human
    Sample Types: Plasma
  7. Persistently raised plasma levels of platelet-derived inflammatory mediators in HIV-infected patients during highly active anti-retroviral therapy.
    Authors: Landro L, Ueland T, Otterdal K, Froland SS, Aukrust P
    J. Thromb. Haemost., 2011;9(5):1075-7.
    Species: Human
    Sample Types: Platelet Poor Plasma
  8. Thrombocytopenia in early malaria is associated with GP1b shedding in absence of systemic platelet activation and consumptive coagulopathy.
    Authors: de Mast Q, de Groot PG, van Heerde WL, Roestenberg M, van Velzen JF, Verbruggen B, Roest M, McCall M, Nieman AE, Westendorp J, Syafruddin D, Fijnheer R, van Dongen-Lases EC, Sauerwein RW, van der Ven AJ
    Br. J. Haematol., 2010;151(5):495-503.
    Species: Human
    Sample Types: Plasma
  9. Systemic inflammatory markers in COPD: results from the Bergen COPD Cohort Study.
    Authors: Eagan TM, Ueland T, Wagner PD, Hardie JA, Mollnes TE, Damas JK, Aukrust P, Bakke PS
    Eur. Respir. J., 2010;35(3):540-8.
    Species: Human
    Sample Types: Plasma
  10. 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;4(11):e7916.
    Species: Human
    Sample Types: Plasma
  11. Connective tissue-activating peptide III: a novel blood biomarker for early lung cancer detection.
    Authors: Yee J, Sadar MD, Sin DD, Kuzyk M, Xing L, Kondra J, McWilliams A, Man SF, Lam S
    J. Clin. Oncol., 2009;27(17):2787-92.
    Species: Human
    Sample Types: Plasma
  12. Increased levels of neutrophil-activating peptide-2 in acute coronary syndromes: possible role of platelet-mediated vascular inflammation.
    Authors: Smith C, Damas JK, Otterdal K, ØIe E, Sandberg WJ, Yndestad A, Waehre T, Scholz H, Endresen K, Olofsson PS, Halvorsen B, Gullestad L, Froland SS, Hansson GK, Aukrust P
    J. Am. Coll. Cardiol., 2006;48(8):1591-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. ELR+ CXC chemokines in human milk.
    Authors: Maheshwari A, Christensen RD, Calhoun DA
    Cytokine, 2003;24(3):91-102.
    Species: Human
    Sample Types: Serum

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