Human CXCL1/GRO alpha Quantikine ELISA Kit, 2nd Gen

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(34 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Specificity
    Natural and recombinant human GRO alpha
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Human CXCL1/GRO alpha Immunoassay is a 4.25-4.5 hour solid phase ELISA designed to measure human GRO alpha in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human GRO alpha and antibodies raised against the recombinant protein. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural human GRO alpha showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human GRO alpha.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean61.425173662.8252734
Standard Deviation2.26203.611.938.9
CV%3.62.42.75.74.75.3

Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean81.431873487.4320758
Standard Deviation3.77.45511.927.662.7
CV%4.52.37.513.68.68.3

Recovery

The recovery of human GRO alpha spiked to levels throughout the range of the assay in various matrices was evaluated.

Linearity
To assess linearity of the assay, samples containing and/or spiked with high concentrations of human GRO alpha were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
    • Entrez Gene IDs
      2919 (Human); 14825 (Mouse); 81503 (Rat);
    • Alternate Names
      CINC1; CINC-1; CXCL1; GRO alpha; KC; MGSA-alpha;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 50 µL Assay Diluent for Serum & Plasma Samples Only
    4.   For Serum & Plasma Samples Only: Add 50 µL of Assay Diluent to each well.

    5. 200 µL Standard, Control, or Sample
    6.   Add 200 µL of Standard, control, or sample to each well.
    7.   For Serum & Plasma Samples: Cover with a plate sealer, and incubate at room temperature for 2 hours.
      For Cell Culture Supernate Samples: Cover with a plate sealer, and incubate at room temperature for 1.5 hours.
    8.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.

    9. 200 µL Conjugate
    10.   Add 200 µL of Conjugate to each well.
    11.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at 2-8 °C for 2 hours.
      For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at 2-8 °C for 1 hour.
    12.   Aspirate and wash 3 times.

    13. 200 µL Substrate Solution
    14. Add 200 µL Substrate Solution to each well.
    15. For Serum & Plasma Samples: Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
      For Cell Culture Supernate Samples: Incubate at room temperature for 15 minutes. PROTECT FROM LIGHT.

    16. 50 µL Stop Solution
    17. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    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.

    34 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. CXCL1 is critical for pre-metastatic niche formation and metastasis in colorectal cancer
      Authors: D Wang, H Sun, J Wei, B Cen, RN DuBois
      Cancer Res., 2017;0(0):.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    2. CAF-secreted CXCL1 conferred radioresistance by regulating DNA damage response in a ROS-dependent manner in esophageal squamous cell carcinoma
      Authors: H Zhang, J Yue, Z Jiang, R Zhou, R Xie, Y Xu, S Wu
      Cell Death Dis, 2017;8(5):e2790.
      Species: Human
      Sample Type: Cell Culture Supernates
    3. p53 and NF-kappaB coregulate proinflammatory gene responses in human macrophages.
      Authors: Lowe J, Menendez D, Bushel P, Shatz M, Kirk E, Troester M, Garantziotis S, Fessler M, Resnick M
      Cancer Res, 2016;74(8):2182-92.
      Species: Human
      Sample Type: Cell Culture Supernates
    4. MicroRNAs regulate cytokine responses in gingival epithelial cells
      Infect Immun, 2016;0(0):.
      Species: Human
      Sample Type: Cell Culture Supernates
    5. High salt primes a specific activation state of macrophages, M(Na).
      Authors: Zhang W, Zheng X, DU L, Sun J, Shen Z, Shi C, Sun S, Zhang Z, Chen X, Qin M, Liu X, Tao J, Jia L, Fan H, Zhou B, Yu Y, Ying H, Hui L, Liu X, Yi X, Liu X, Zhang L, Duan S
      Cell Res, 2015;25(8):893-910.
      Species: Human
      Sample Type: Cell Culture Supernates
    6. Stromal cells positively and negatively modulate the growth of cancer cells: stimulation via the PGE2-TNFalpha-IL-6 pathway and inhibition via secreted GAPDH-E-cadherin interaction.
      Authors: Kawada M, Inoue H, Ohba S, Yoshida J, Masuda T, Yamasaki M, Usami I, Sakamoto S, Abe H, Watanabe T, Yamori T, Shibasaki M, Nomoto A
      PLoS ONE, 2015;10(3):e0119415.
      Species: Human
      Sample Type: Cell Culture Supernates
    7. Phosphoinositide 3-kinase inhibition restores neutrophil accuracy in the elderly: toward targeted treatments for immunosenescence.
      Authors: Sapey E, Greenwood H, Walton G, Mann E, Love A, Aaronson N, Insall R, Stockley R, Lord J
      Blood, 2014;123(2):239-48.
      Species: Human
      Sample Type: Sputum
    8. Circulating levels of transforming growth factor-betaeta (TGF-beta) and chemokine (C-X-C motif) ligand-1 (CXCL1) as predictors of distant seeding of circulating tumor cells in patients with metastatic breast cancer.
      Authors: Divella R, Daniele A, Savino E, Palma F, Bellizzi A, Giotta F, Simone G, Lioce M, Quaranta M, Paradiso A, Mazzocca A
      Anticancer Res, 2013;33(4):1491-7.
      Species: Human
      Sample Type: Plasma
    9. Targeted gene silencing using a follicle-stimulating hormone peptide-conjugated nanoparticle system improves its specificity and efficacy in ovarian clear cell carcinoma in vitro.
      Authors: Hong S, Zhang X, Chen J, Zhou J, Zheng Y, Xu C
      J Ovarian Res, 2013;6(1):80.
      Species: Human
      Sample Type: Cell Culture Supernates
    10. Influence of the CXCL1 rs4074 A allele on alcohol induced cirrhosis and HCC in patients of European descent.
      Authors: Nischalke H, Berger C, Lutz P, Langhans B, Wolter F, Eisenhardt M, Kramer B, Kokordelis P, Glassner A, Muller T, Rosendahl J, Fischer J, Berg T, Grunhage F, Leifeld L, Soyka M, Nattermann J, Sauerbruch T, Stickel F, Spengler U
      PLoS ONE, 2013;8(11):e80848.
      Species: Human
      Sample Type: Cell Culture Supernates
    11. Role of interleukin-32 in Helicobacter pylori-induced gastric inflammation.
      Authors: Sakitani K, Hirata Y, Hayakawa Y, Serizawa T, Nakata W, Takahashi R, Kinoshita H, Sakamoto K, Nakagawa H, Akanuma M, Yoshida H, Maeda S, Koike K
      Infect Immun, 2012;80(11):3795-803.
      Species: Human
      Sample Type: Cell Lysates
    12. Thymic stromal lymphopoietin induces chemotactic and prosurvival effects in eosinophils: implications in allergic inflammation.
      Authors: Wong CK, Hu S, Cheung PF
      Am. J. Respir. Cell Mol. Biol., 2010;43(3):305-15.
      Species: Human
      Sample Type: Cell Culture Supernates
    13. CXCR2 promotes ovarian cancer growth through dysregulated cell cycle, diminished apoptosis, and enhanced angiogenesis.
      Authors: Yang G, Rosen DG, Liu G
      Clin. Cancer Res., 2010;16(15):3875-86.
      Species: Human
      Sample Type: Cell Culture Supernates
    14. A phase I trial of bortezomib with temozolomide in patients with advanced melanoma: toxicities, antitumor effects, and modulation of therapeutic targets.
      Authors: Su Y, Amiri KI, Horton LW
      Clin. Cancer Res., 2010;16(1):348-57.
      Species: Human
      Sample Type: Serum
    15. Immunomodulation of nasal epithelial cells by Staphylococcus aureus-derived serine proteases.
      Authors: Rudack C, Sachse F, Albert N, Becker K, von Eiff C
      J. Immunol., 2009;183(11):7592-601.
      Species: Human
      Sample Type: Cell Culture Supernates
    16. LPS induces GROalpha chemokine production via NF-kappaB in oral fibroblasts.
      Authors: Jonsson D, Amisten S, Bratthall G, Holm A, Nilsson BO
      Inflamm. Res., 2009;58(11):791-6.
      Species: Human
      Sample Type: Cell Culture Supernates
    17. Dysregulation of stromal derived factor 1/CXCR4 axis in the megakaryocytic lineage in essential thrombocythemia.
      Authors: Salim JP, Goette NP, Lev PR, Chazarreta CD, Heller PG, Alvarez C, Molinas FC, Marta RF
      Br. J. Haematol., 2009;144(1):69-77.
      Species: Human
      Sample Type: Plasma
    18. Duffy antigen modifies the chemokine response in human endotoxemia.
      Authors: Mayr FB, Spiel AO, Leitner JM, Firbas C, Kliegel T, Jilma-Stohlawetz P, Derendorf H, Jilma B
      Crit. Care Med., 2008;36(1):159-65.
      Species: Human
      Sample Type: Plasma
    19. MMP-12 induces IL-8/CXCL8 secretion through EGFR and ERK1/2 activation in epithelial cells.
      Authors: Le Quement C, Guenon I, Gillon JY, Lagente V, Boichot E
      Am. J. Physiol. Lung Cell Mol. Physiol., 2008;294(6):L1076-84.
      Species: Human
      Sample Type: Cell Culture Supernates
    20. CT20126, a novel immunosuppressant, prevents collagen-induced arthritis through the down-regulation of inflammatory gene expression by inhibiting NF-kappaB activation.
      Authors: Lee SJ, Nam WD, Na HJ, Cho YL, Ha KS, Hwang JY, Lee H, Kim SO, Kwon YG, Kim YM
      Biochem. Pharmacol., 2008;76(1):79-90.
      Species: Human
      Sample Type: Cell Culture Supernates
    21. IL-8 as antibody therapeutic target in inflammatory diseases: reduction of clinical activity in palmoplantar pustulosis.
      Authors: Skov L, Beurskens FJ, Zachariae CO, Reitamo S, Teeling J, Satijn D, Knudsen KM, Boot EP, Hudson D, Baadsgaard O, Parren PW, van de Winkel JG
      J. Immunol., 2008;181(1):669-79.
      Species: Human
      Sample Type: Complex Sample Type
    22. A potential role of the CXC chemokine GROalpha in atherosclerosis and plaque destabilization: downregulatory effects of statins.
      Authors: Breland UM, Halvorsen B, Hol J, ØIe E, Paulsson-Berne G, Yndestad A, Smith C, Otterdal K, Hedin U, Waehre T, Sandberg WJ, Froland SS, Haraldsen G, Gullestad L, Damas JK, Hansson GK, Aukrust P
      Arterioscler. Thromb. Vasc. Biol., 2008;28(5):1005-11.
      Species: Human
      Sample Type: Plasma
    23. The human IL-17F/IL-17A heterodimeric cytokine signals through the IL-17RA/IL-17RC receptor complex.
      Authors: Wright JF, Bennett F, Li B, Brooks J, Luxenberg DP, Whitters MJ, Tomkinson KN, Fitz LJ, Wolfman NM, Collins M, Dunussi-Joannopoulos K, Chatterjee-Kishore M, Carreno BM
      J. Immunol., 2008;181(4):2799-805.
      Species: Human
      Sample Type: Cell Culture Supernates
    24. Role of interleukin-8 and growth-regulated oncogene-alpha in the chemotactic migration of all-trans retinoic acid-treated promyelocytic leukemic cells toward alveolar epithelial cells.
      Authors: Tsai WH, Hsu HC, Lin CC, Ho CK, Kou YR
      Crit. Care Med., 2007;35(3):879-85.
      Species: Human
      Sample Type: Cell Culture Supernates
    25. Proliferation, differentiation, and cytokine secretion of human umbilical cord blood-derived mononuclear cells in vitro.
      Authors: Neuhoff S, Moers J, Rieks M, Grunwald T, Jensen A, Dermietzel R, Meier C
      Exp. Hematol., 2007;35(7):1119-31.
      Species: Human
      Sample Type: Cell Culture Supernates
    26. Th1/Th2 cytokines reciprocally regulate in vitro pulmonary angiogenesis via CXC chemokine synthesis.
      Authors: Matsuda A, Fukuda S, Matsumoto K, Saito H
      Am. J. Respir. Cell Mol. Biol., 2007;38(2):168-75.
      Species: Human
      Sample Type: Cell Culture Supernates
    27. L5, the most electronegative subfraction of plasma LDL, induces endothelial vascular cell adhesion molecule 1 and CXC chemokines, which mediate mononuclear leukocyte adhesion.
      Authors: Abe Y, Fornage M, Yang CY, Bui-Thanh NA, Wise V, Chen HH, Rangaraj G, Ballantyne CM
      Atherosclerosis, 2007;192(1):56-66.
      Species: Human
      Sample Type: Cell Culture Supernates
    28. Different angiogenic activity in pulmonary sarcoidosis and idiopathic pulmonary fibrosis.
      Authors: Antoniou KM, Tzouvelekis A, Alexandrakis MG, Sfiridaki K, Tsiligianni I, Rachiotis G, Tzanakis N, Bouros D, Milic-Emili J, Siafakas NM
      Chest, 2006;130(4):982-8.
      Species: Human
      Sample Type: BALF
    29. Exhaled breath condensate levels of 8-isoprostane, growth related oncogene alpha and monocyte chemoattractant protein-1 in patients with chronic obstructive pulmonary disease.
      Authors: Ko FW, Lau CY, Leung TF, Wong GW, Lam CW, Hui DS
      Respir Med, 2006;100(4):630-8.
      Species: Human
      Sample Type: Exhaled Breath Condensate (EBC)
    30. IL-17 enhances the net angiogenic activity and in vivo growth of human non-small cell lung cancer in SCID mice through promoting CXCR-2-dependent angiogenesis.
      Authors: Numasaki M, Watanabe M, Suzuki T, Takahashi H, Nakamura A, McAllister F, Hishinuma T, Goto J, Lotze MT, Kolls JK, Sasaki H
      J. Immunol., 2005;175(9):6177-89.
      Species: Human
      Sample Type: Cell Culture Supernates
    31. ELR+ CXC chemokines in human milk.
      Authors: Maheshwari A, Christensen RD, Calhoun DA
      Cytokine, 2003;24(3):91-102.
      Species: Human
      Sample Type: Serum
    32. TNFalpha-induced macrophage chemokine secretion is more dependent on NF-kappaB expression than lipopolysaccharides-induced macrophage chemokine secretion.
      Authors: Ciesielski CJ, Andreakos E, Foxwell BM, Feldmann M
      Eur. J. Immunol., 2002;32(7):2037-45.
      Species: Human
      Sample Type: Cell Culture Supernates
    33. Il-6 and its soluble receptor orchestrate a temporal switch in the pattern of leukocyte recruitment seen during acute inflammation.
      Authors: Hurst SM, Wilkinson TS, McLoughlin RM, Jones S, Horiuchi S, Yamamoto N, Rose-John S, Fuller GM, Topley N, Jones SA
      Immunity, 2001;14(6):705-14.
      Species: Human
      Sample Type: Cell Culture Supernates
    34. High EMT Signature Score of Invasive Non-Small Cell Lung Cancer (NSCLC) Cells Correlates with NFkappaB Driven Colony-Stimulating Factor 2 (CSF2/GM-CSF) Secretion by Neighboring Stromal Fibroblasts.
      Authors: Rudisch A, Dewhurst M, Horga L, Kramer N, Harrer N, Dong M, van der Kuip H, Wernitznig A, Bernthaler A, Dolznig H, Sommergruber W
      PLoS ONE, 0;10(4):e0124283.
      Species: Human
      Sample Type: Cell Culture Supernates
    Expand to show all 34 Citations

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