Human CXCL1/GRO alpha 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 CXCL1/GROalpha. 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)
Data Example
Product Datasheets
Preparation and Storage
Background: CXCL1/GRO alpha/KC/CINC-1
CXCL1/GRO alpha, also known as KC in mouse and CINC-1 in rat, is a member of the CXC family of chemokines. Human CXCL1/GRO alpha is 107 amino acids (aa) in length with a predicted molecular weight of 11 kDa. The mouse and rat orthologs share 57% and 59% amino acid sequence identity with the human protein, respectively. Genome-wide analysis suggests that the protein encoded by the mouse Cxcl3 gene is homologous to human CXCL1/GRO alpha protein. Human CXCL1/GRO alpha can be cleaved into three isoforms CXCL1/GRO alpha (aa 4-73), CXCL1/GRO alpha (aa 5-73), and CXCL1/GRO alpha (aa 6-73) which show higher activity than the full length protein. CXCL1/GRO alpha is secreted by mast cells and macrophages whereupon it acts as a chemoattractant for 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 CXCL1/GRO alpha 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.
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Citations: Showing 1 - 10
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Constitutive Interferon Attenuates RIPK1/3-Mediated Cytokine Translation
Authors: HI Muendlein, J Sarhan, BC Liu, WM Connolly, SA Schworer, I Smirnova, AY Tang, V Ilyukha, J Pietruska, S Tahmasebi, N Sonenberg, A Degterev, A Poltorak
Cell Rep, 2020;30(3):699-713.e4.
Species: Human
Sample Types: Cell Culture Media
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Bleomycin hydrolase regulates the release of chemokines important for inflammation and wound healing by keratinocytes
Authors: R Riise, L Odqvist, J Mattsson, S Monkley, SM Abdillahi, C Tyrchan, D Muthas, LF Yrlid
Sci Rep, 2019;9(1):20407.
Species: Human
Sample Types: Cell Culture Supernates
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Reduced expression of miR-146a in human bronchial epithelial cells alters neutrophil migration
Authors: A Kivihall, A Aab, J Soja, K S?adek, M Sanak, A Altraja, B Jakiela, G Bochenek, A Rebane
Clin Transl Allergy, 2019;9(0):62.
Species: Human
Sample Types: Cell Culture Supernates
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Regulation of CXCL1 chemokine and CSF3 cytokine levels in myometrial cells by the MAFF transcription factor
Authors: J Saliba, B Coutaud, V Solovieva, F Lu, V Blank
J. Cell. Mol. Med., 2019;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Rhinovirus and Bacteria Synergistically Induce IL-17C Release from Human Airway Epithelial Cells To Promote Neutrophil Recruitment
Authors: KC Jamieson, SL Traves, C Kooi, S Wiehler, CJ Dumonceaux, BA Maciejewsk, JW Arnason, AN Michi, R Leigh, D Proud
J. Immunol., 2018;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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A bispecific antibody that targets IL-6 receptor and IL-17A for the potential therapy of patients with autoimmune and inflammatory diseases
Authors: M Lyman, V Lieuw, R Richardson, A Timmer, C Stewart, S Granger, R Woods, M Silacci, D Grabulovsk, R Newman
J. Biol. Chem., 2018;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Placental Ras Regulates Inflammation Associated with Maternal Obesity
Authors: S Liong, G Barker, M Lappas
Mediators Inflamm., 2018;2018(0):3645386.
Species: Human
Sample Types: Cell Culture Supernates
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Neutrophil recruitment to inflamed joints can occur without cellular priming
Authors: L Björkman, K Christenso, L Davidsson, J Mårtensson, F Amirbeagi, A Welin, H Forsman, A Karlsson, C Dahlgren, J Bylund
J. Leukoc. Biol., 2018;0(0):.
Species: Human
Sample Types: Serum
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The small molecule rhodomyrtone suppresses TNF-? and IL-17A-induced keratinocyte inflammatory responses: A potential new therapeutic for psoriasis
Authors: J Chorachoo, S Lambert, T Furnholm, L Roberts, L Reingold, S Auepemkiat, SP Voravuthik, A Johnston
PLoS ONE, 2018;13(10):e0205340.
Species: Human
Sample Types: Cell Culture Supernates
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Inhibition of IRE1 RNase activity modulates the tumor cell secretome and enhances response to chemotherapy
Authors: SE Logue, EP McGrath, P Cleary, S Greene, K Mnich, A Almanza, E Chevet, RM Dwyer, A Oommen, P Legembre, F Godey, EC Madden, B Leuzzi, J Obacz, Q Zeng, JB Patterson, R Jäger, AM Gorman, A Samali
Nat Commun, 2018;9(1):3267.
Species: Human
Sample Types: Cell Culture Supernates
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The TRAIL-Induced Cancer Secretome Promotes a Tumor-Supportive Immune Microenvironment via CCR2
Authors: T Hartwig, A Montinaro, S von Karste, A Sevko, S Surinova, A Chakravart, L Taraborrel, P Draber, E Lafont, F Arce Varga, MA El-Bahrawy, SA Quezada, H Walczak
Mol. Cell, 2017;65(4):730-742.e5.
Species: Human
Sample Types: Cell Culture Supernates
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Caspase-8 Acts in a Non-enzymatic Role as a Scaffold for Assembly of a Pro-inflammatory FADDosome Complex upon TRAIL Stimulation
Authors: CM Henry, SJ Martin
Mol. Cell, 2017;65(4):715-729.e5.
Species: Human
Sample Types: Cell Culture Supernates
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Syndecan-1 Acts as an Important Regulator of CXCL1 Expression and Cellular Interaction of Human Endometrial Stromal and Trophoblast Cells
Authors: DM Baston-Bue, O Altergot-A, SJ Pour, JS Krüssel, UR Markert, TN Fehm, AP Bielfeld
Mediators Inflamm., 2017;2017(0):8379256.
Species: Human
Sample Types: Cell Culture Supernates
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The Proangiogenic Capabilities of Malignant Ascites Generated by Aggressive Ovarian Tumors
Authors: J Miku?a-Pie, P Uruski, S Szubert, K Maksin, R Moszy?ski, D Szpurek, A Wo?niak, S Sajdak, A Tykarski, K Ksi??ek
Biomed Res Int, 2017;2017(0):2592496.
Species: Human
Sample Types: Cell Culture Supernates
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Pathogenic role of glycan-specific IgG antibodies in IgA nephropathy
Authors: YF Zhao, L Zhu, LJ Liu, SF Shi, JC Lv, H Zhang
BMC Nephrol, 2017;18(1):301.
Species: Human
Sample Types: Cell Culture Supernates
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Serum from Varicose Patients Induces Senescence-Related Dysfunction of Vascular Endothelium Generating Local and Systemic Proinflammatory Conditions
Oxid Med Cell Longev, 2016;2016(0):2069290.
Species: Human
Sample Types: Serum
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Inhibiting complex IL-17A and IL-17RA interactions with a linear peptide
Sci Rep, 2016;6(0):26071.
Species: Human
Sample Types: Cell Culture Supernates
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Macrophage activation in acute exacerbation of idiopathic pulmonary fibrosis.
Authors: Schupp J, Binder H, Jager B, Cillis G, Zissel G, Muller-Quernheim J, Prasse A
PLoS ONE, 2015;10(1):e0116775.
Species: Human
Sample Types: BALF
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Necroptosis suppresses inflammation via termination of TNF- or LPS-induced cytokine and chemokine production.
Authors: Kearney C, Cullen S, Tynan G, Henry C, Clancy D, Lavelle E, Martin S
Cell Death Differ, 2015;22(8):1313-27.
Species: Human
Sample Types: Cell Culture Supernates
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Urinary CXCL1: a novel predictor of IgA nephropathy progression.
Authors: Zhao, Yanfeng, Zhu, Li, Zhou, Tong, Zhang, Qingxian, Shi, Sufang, Liu, Lijun, Lv, Jicheng, Zhang, Hong
PLoS ONE, 2015;10(3):e0119033.
Species: Human
Sample Types: Urine
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Human osteoarthritic cartilage shows reduced in vivo expression of IL-4, a chondroprotective cytokine that differentially modulates IL-1beta-stimulated production of chemokines and matrix-degrading enzymes in vitro.
Authors: Assirelli E, Pulsatelli L, Dolzani P, Platano D, Olivotto E, Filardo G, Trisolino G, Facchini A, Borzi R, Meliconi R
PLoS ONE, 2014;9(5):e96925.
Species: Human
Sample Types: Cell Culture Supernates
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TNF-alpha induces CXCL1 chemokine expression and release in human vascular endothelial cells in vitro via two distinct signaling pathways.
Authors: Lo H, Lai T, Li C, Wu W
Acta Pharmacol Sin, 2014;35(3):339-50.
Species: Human
Sample Types: Cell Culture Supernates
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Roles for the mitogen-activated protein kinase (MAPK) phosphatase, DUSP1, in feedback control of inflammatory gene expression and repression by dexamethasone.
Authors: Shah S, King E, Chandrasekhar A, Newton R
J Biol Chem, 2014;289(19):13667-79.
Species: Human
Sample Types: Cell Culture Supernates
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Chinese herbal medicine suppresses invasion-promoting capacity of cancer-associated fibroblasts in pancreatic cancer.
Authors: Chen, Lianyu, Qu, Chao, Chen, Hao, Xu, Litao, Qi, Qi, Luo, Jianmin, Wang, Kun, Meng, Zhiqiang, Chen, Zhen, Wang, Peng, Liu, Luming
PLoS ONE, 2014;9(4):e96177.
Species: Human
Sample Types: Cell Culture Supernates
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Selective activation of human dendritic cells by OM-85 through a NF-kB and MAPK dependent pathway.
Authors: Parola C, Salogni L, Vaira X, Scutera S, Somma P, Salvi V, Musso T, Tabbia G, Bardessono M, Pasquali C, Mantovani A, Sozzani S, Bosisio D
PLoS ONE, 2013;8(12):e82867.
Species: Human
Sample Types: Cell Culture Supernates
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Serglycin is a major proteoglycan in polarized human endothelial cells and is implicated in the secretion of the chemokine GROalpha/CXCL1.
Authors: Meen AJ, Oynebraten I, Reine TM, Duelli A, Svennevig K, Pejler G, Jenssen T, Kolset SO
J. Biol. Chem., 2011;286(4):2636-47.
Species: Human
Sample Types: Cell Lysates
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Chronic mucocutaneous candidiasis in APECED or thymoma patients correlates with autoimmunity to Th17-associated cytokines.
Authors: Kisand K, Boe Wolff AS, Podkrajsek KT, Tserel L, Link M, Kisand KV, Ersvaer E, Perheentupa J, Erichsen MM, Bratanic N, Meloni A, Cetani F, Perniola R, Ergun-Longmire B, Maclaren N, Krohn KJ, Pura M, Schalke B, Strobel P, Leite MI, Battelino T, Husebye ES, Peterson P, Willcox N, Meager A
J. Exp. Med., 2010;207(2):299-308.
Species: Human
Sample Types: Cell Culture Supernates
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Impact of tobacco-smoke on key signaling pathways in the innate immune response in lung macrophages.
Authors: Birrell MA, Wong S, Catley MC, Belvisi MG
J. Cell. Physiol., 2008;214(1):27-37.
Species: Human
Sample Types: Cell Culture Supernates
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The inflammatory microenvironment of the aging prostate facilitates cellular proliferation and hypertrophy.
Authors: Begley LA, Kasina S, Macdonald J, Macoska JA
Cytokine, 2008;43(2):194-9.
Species: Human
Sample Types: Cell Culture Supernates
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Novel role for the liver X nuclear receptor in the suppression of lung inflammatory responses.
Authors: Birrell MA, Catley MC, Hardaker E, Wong S, Willson TM, McCluskie K, Leonard T, Farrow SN, Collins JL, Haj-Yahia S, Belvisi MG
J. Biol. Chem., 2007;282(44):31882-90.
Species: Human
Sample Types: Cell Culture Supernates
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Increased ENA-78 in the follicular fluid of patients with endometriosis.
Authors: Wunder DM, Mueller MD, Birkhauser MH, Bersinger NA
Acta Obstet Gynecol Scand, 2006;85(3):336-42.
Species: Human
Sample Types: Follicular Fluid
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Role of matrix metalloproteinases in the inflammatory response in human airway cell-based assays and in rodent models of airway disease.
Authors: Birrell MA, Wong S, Dekkak A, De Alba J, Haj-Yahia S, Belvisi MG
J. Pharmacol. Exp. Ther., 2006;318(2):741-50.
Species: Human
Sample Types: Cell Culture Supernates
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Rapid chemokine secretion from endothelial cells originates from 2 distinct compartments.
Authors: Oynebraten I, Bakke O, Brandtzaeg P, Johansen FE, Haraldsen G
Blood, 2004;104(2):314-20.
Species: Human
Sample Types: Cell Culture Supernates
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Interleukin (IL)-4, IL-13, and IL-17A differentially affect the profibrotic and proinflammatory functions of fibrocytes from asthmatic patients.
Authors: Bellini A, Marini MA, Bianchetti L, Barczyk M, Schmidt M, Mattoli S
Mucosal Immunol, 0;5(2):140-9.
Species: Human
Sample Types: Cell Culture Supernates
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