Human CXCL2/GRO beta 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.
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: CXCL2/GRO beta/MIP-2/CINC-3
CXCL2/GRO beta, also called MIP-2 in mouse and CINC-3 in rat, is a member of the CXC chemokine family. Human CXCL2/GRO beta is 107 amino acids (aa) in length with a predicted molecular weight of 11 kDa. The mouse and rat orthologs share 70% and 71% aa sequence identity with the human protein, respectively. N-terminal aa 1-4 of CXCL2/GRO beta can be post-translationally cleaved which confers enhanced hematopoietic bioactivity. CXCL2/GRO beta is produced by a variety of cell types including monocytes and macrophages at sites of inflammation and is chemotactic for granulocytes, including neutrophils.
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 CXCL2/GRO beta 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.
12
Citations: Showing 1 - 10
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Serum interleukin-6, procalcitonin, and C-reactive protein at hospital admission can identify patients at low risk for severe COVID-19 progression
Authors: Zobel, CM;Wenzel, W;Krüger, JP;Baumgarten, U;Wagelöhner, T;Neumann, N;Foroutan, B;Müller, R;Müller, A;Rauschning, D;Schü beta ler, M;Scheit, L;Weinreich, F;Oltmanns, K;Keidel, F;Koch, M;Spethmann, S;Schreiner, M;
Frontiers in microbiology
Species: Human
Sample Types: Serum
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c-Met Mediated Cytokine Network Promotes Brain Metastasis of Breast Cancer by Remodeling Neutrophil Activities
Authors: Liu, Y;Smith, MR;Wang, Y;D'Agostino, R;Ruiz, J;Lycan, T;Kucera, GL;Miller, LD;Li, W;Chan, MD;Farris, M;Su, J;Song, Q;Zhao, D;Chandrasekaran, A;Xing, F;
Cancers
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KDM6A Loss Recruits Tumor-Associated Neutrophils and Promotes Neutrophil Extracellular Trap Formation in Pancreatic Cancer
Authors: J Yang, L Jin, HS Kim, F Tian, Z Yi, K Bedi, M Ljungman, M Pasca di M, H Crawford, J Shi
Cancer Research, 2022-11-15;0(0):OF1-OF14.
Species: Human
Sample Types: Cell Culture Supernates
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Multi-level inhibition of coronavirus replication by chemical ER stress
Authors: MS Shaban, C Müller, C Mayr-Buro, H Weiser, J Meier-Soel, BV Albert, A Weber, U Linne, T Hain, I Babayev, N Karl, N Hofmann, S Becker, S Herold, ML Schmitz, J Ziebuhr, M Kracht
Nature Communications, 2021-09-20;12(1):5536.
Species: Human
Sample Types: Cell Culture Supernates
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Patient Derived Colonoids as Drug Testing Platforms-Critical Importance of Oxygen Concentration
Authors: HK Skovdahl, S Gopalakris, TD Svendsen, AVB Granlund, I Bakke, ZG Ginbot, S Thorsvik, A Flatberg, B Sporsheim, J Ostrop, TE Mollnes, AK Sandvik, T Bruland
Frontiers in Pharmacology, 2021-05-13;12(0):679741.
Species: Human
Sample Types: Cell Culture Supernates
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Macrophage development and activation involve coordinated intron retention in key inflammatory regulators
Authors: ID Green, N Pinello, R Song, Q Lee, JM Halstead, CT Kwok, ACH Wong, SS Nair, SJ Clark, B Roediger, U Schmitz, M Larance, R Hayashi, JEJ Rasko, JJ Wong
Nucleic Acids Res., 2020-07-09;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2019-10-07;10(10):760.
Species: Human
Sample Types: Cell Culture Supernates
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Resistance to lysosomotropic drugs used to treat kidney and breast cancers involves autophagy and inflammation and converges in inducing CXCL5
Authors: S Giuliano, M Dufies, PD Ndiaye, J Viotti, D Borchielli, J Parola, V Vial, Y Cormerais, M Ohanna, V Imbert, E Chamorey, N Rioux-Lecl, A Savina, JM Ferrero, B Mograbi, G Pagès
Theranostics, 2019-01-30;9(4):1181-1199.
Species: Human
Sample Types: Cell Culture Supernates
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Fluid restriction reduces pulmonary edema in a model of acute lung injury in mechanically ventilated rats
Authors: SA Ingelse, J Juschten, MAW Maas, G Matute-Bel, NP Juffermans, JBM van Woense, RA Bem
PLoS ONE, 2019-01-17;14(1):e0210172.
Species: Human
Sample Types: Tissue Homogenates
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Transporters MRP1 and MRP2 Regulate Opposing Inflammatory Signals To Control Transepithelial Neutrophil Migration during Streptococcus pneumoniae Lung Infection
Authors: A Zukauskas, RJ Mrsny, P Cortés Bar, JR Turner, JM Leong, BA McCormick
mSphere, 2018-07-05;3(4):.
Species: Human
Sample Types: Cell Culture Supernates
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Intracapillary immune complexes recruit and activate slan-expressing CD16+ monocytes in human lupus nephritis
Authors: F Olaru, T Döbel, AS Lonsdorf, S Oehrl, M Maas, AH Enk, M Schmitz, EF Gröne, HJ Gröne, K Schäkel
JCI Insight, 2018-06-07;3(11):.
Species: Human
Sample Types: Cell Culture Supernates
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The Fli-1 transcription factor is a critical regulator for controlling the expression of chemokine C-X-C motif ligand 2 (CXCL2)
Authors: Xian K Zhang
Mol. Immunol., 2016-11-24;81(0):59-66.
Species: Human
Sample Types: Cell Culture Supernates
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