Human CXCL5/ENA-78 Quantikine ELISA Kit

  (21 citations)
(1 Review)
    
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Assay Procedure
Citations (21)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (10 uL), Serum (25 uL), EDTA Plasma (50 uL), Platelet-poor Heparin Plasma (50 uL), Citrate Plasma (50 uL)
  • Sensitivity
    15 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Platelet-poor Heparin Plasma, Citrate Plasma)
  • Specificity
    Natural and recombinant human ENA-78
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC22 , Quantikine Immunoassay Control Group 7 - Please Inquire
Product Summary
The Quantikine Human ENA-78 Immunoassay is a 4.5 hour solid phase ELISA designed to measure human ENA-78 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human ENA-78 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant human ENA-78 accurately. Results obtained using natural human ENA-78 showed dose curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human ENA-78.

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 separate assays to assess inter-assay precision
Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 112 251 1096 129 278 1085
Standard Deviation 6 14.6 44.9 12.6 26.8 96.5
CV% 5.4 5.8 4.1 9.8 9.6 8.9

Serum, EDTA Plasma, Platelet-poor Heparin Plasma, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 113 244 995 109 240 989
Standard Deviation 9.4 12.9 37.8 10.1 17.8 65.8
CV% 8.3 5.3 3.8 9.3 7.4 6.7

Recovery

The recovery of ENA-78 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 101 95-107
Citrate Plasma (n=5) 100 94-106
EDTA Plasma (n=5) 98 92-104
Heparin Plasma (n=5) 98 93-103
Serum (n=5) 101 93-109
Linearity
To assess the linearity of the assay, five samples containing or spiked with high concentrations of ENA-78 in various matrices were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
 CXCL5/ENA-78 [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CXCL5/ENA-78
CXCL5/Epithelial Cell-derived Neutrophil-activating Peptide (ENA-78), is a member of the CXC subfamily of chemokines. Full-length CXCL5/ENA-78 is 114 amino acids (aa) in length with a predicted molecular weight of 12 kDa. Following the removal of the signal peptide, bioactive CXCL5/ENA-78 is 78 aa in length. CXCL5/ENA-78 can be N-terminally cleaved by Cathepsin G and Chymotrypsin to CXCL5/ENA-74 (74 aa) and CXCL5/ENA-70 (70 aa), which show increased potency relative to CXCL5/ENA-78. While murine LIX was thought to be an ortholog to human CXCL5/ENA-78, genome-wide analysis and a consensus in the field suggests that human CXCL5/ENA-78 does not have a true murine ortholog. CXCL5/ENA-78 is upregulated at sites of inflammation and is expressed by multiple hematopoietic cell types, fibroblasts, endothelial cells, vascular smooth muscle cells, and adipocytes. It acts as a chemoattractant for neutrophils, has angiogenic properties, and contributes to cancer progression.
  • Entrez Gene IDs:
    6374 (Human)
  • Alternate Names:
    CXCL5; CXCL5/ENA-78; ENA78; ENA-78
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. 200 µL Assay Diluent
  4.   Add 200 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 3 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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.

21 Citations: Showing 1 - 10
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Species
Sample Type
  1. The CXCL5/CXCR2 axis contributes to the epithelial-mesenchymal transition of nasopharyngeal carcinoma cells by activating ERK/GSK-3?/snail signalling
    Authors: WZ Qiu, HB Zhang, WX Xia, LR Ke, J Yang, YH Yu, H Liang, XJ Huang, GY Liu, WZ Li, YQ Xiang, TB Kang, X Guo, X Lv
    J. Exp. Clin. Cancer Res., 2018;37(1):85.
    Species: Human
    Sample Type: Serum
  2. Caspase 3 involves in neuroplasticity, microglial activation and neurogenesis in the mice hippocampus after intracerebral injection of kainic acid.
    Authors: Tzeng, Tsai-Ten, Tsay, Huey-Jen, Chang, Luping, Hsu, Chia-Lin, Lai, Tzu-Hsua, Huang, Fong-Lee, Shiao, Young-Ji
    J Biomed Sci, 2013;20(0):90.
    Species: Human
    Sample Type: Cell Culture Supernates
  3. Decreased plasma cytokines are associated with low platelet counts in aplastic anemia and immune thrombocytopenic purpura.
    J. Thromb. Haemost., 2012;10(8):1616-23.
    Species: Human
    Sample Type: Serum
  4. Inhibition of IL-1beta-mediated inflammatory responses by the IkappaB alpha super-repressor in human fibroblast-like synoviocytes.
    Authors: Lee YR, Kweon SH, Kwon KB, Park JW, Yoon TR, Park BH
    Biochem. Biophys. Res. Commun., 2009;378(1):90-4.
    Species: Human
    Sample Type: Cell Culture Supernates
  5. Guggulsterone blocks IL-1beta-mediated inflammatory responses by suppressing NF-kappaB activation in fibroblast-like synoviocytes.
    Authors: Lee YR, Lee JH, Noh EM, Kim EK, Song MY, Jung WS, Park SJ, Kim JS, Park JW, Kwon KB, Park BH
    Life Sci., 2008;82(23):1203-9.
    Species: Human
    Sample Type: Cell Culture Supernates
  6. 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
  7. Intestinal epithelial cell up-regulation of LY6 molecules during colitis results in enhanced chemokine secretion.
    Authors: Flanagan K, Modrusan Z, Cornelius J, Chavali A, Kasman I, Komuves L, Mo L, Diehl L
    J. Immunol., 2008;180(6):3874-81.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  8. 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
  9. Coevolution of prostate cancer and bone stroma in three-dimensional coculture: implications for cancer growth and metastasis.
    Authors: Sung SY, Hsieh CL, Law A, Zhau HE, Pathak S, Multani AS, Lim S, Coleman IM, Wu LC, Figg WD, Dahut WL, Nelson P, Lee JK, Amin MB, Lyles R, Johnstone PA, Marshall FF, Chung LW
    Cancer Res., 2008;68(23):9996-10003.
    Species: Human
    Sample Type: Serum
  10. 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
  11. Cooperative effects of rhinovirus and TNF-{alpha} on airway epithelial cell chemokine expression.
    Authors: Newcomb DC, Sajjan US, Nagarkar DR, Goldsmith AM, Bentley JK, Hershenson MB
    Am. J. Physiol. Lung Cell Mol. Physiol., 2007;293(4):L1021-8.
    Species: Human
    Sample Type: Cell Culture Supernates
  12. Crucial role of the protein C pathway in governing microvascular inflammation in inflammatory bowel disease.
    Authors: Scaldaferri F, Sans M, Vetrano S, Graziani C, De Cristofaro R, Gerlitz B, Repici A, Arena V, Malesci A, Panes J, Grinnell BW, Danese S
    J. Clin. Invest., 2007;117(7):1951-60.
    Species: Human
    Sample Type: Cell Culture Supernates
  13. 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
  14. Expression and gene polymorphisms of the chemokine CXCL5 in colorectal cancer patients.
    Authors: Dimberg J, Dienus O, Lofgren S, Hugander A, Wagsater D
    Int. J. Oncol., 2007;31(1):97-102.
    Species: Human
    Sample Type: Plasma
  15. 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
  16. 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
  17. Peritoneal fluid concentrations of epithelial neutrophil-activating peptide-78 correlate with the severity of endometriosis.
    Authors: Suzumori N, Katano K, Suzumori K
    Fertil. Steril., 2004;81(2):305-8.
    Species: Human
    Sample Type: Peritoneal Fluid
  18. CXC-chemokine stimulation of neutrophils correlates with plasma levels of myeloperoxidase and lactoferrin and contributes to clinical outcome after pediatric cardiac surgery.
    Authors: Gessler P, Pretre R, Hohl V, Rousson V, Fischer J, Dahinden C
    Shock, 2004;22(6):513-20.
    Species: Human
    Sample Type: Plasma
  19. ELR+ CXC chemokines in human milk.
    Authors: Maheshwari A, Christensen RD, Calhoun DA
    Cytokine, 2003;24(3):91-102.
    Species: Human
    Sample Type: Serum
  20. Evasion from NK cell immunity by MHC class I chain-related molecules expressing colon adenocarcinoma.
    Authors: Doubrovina ES, Doubrovin MM, Vider E, Sisson RB, O&amp;amp;apos;Reilly RJ, Dupont B, Vyas YM
    J. Immunol., 2003;171(12):6891-9.
    Species: Human
    Sample Type: Serum
  21. 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
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Average Rating: 5 (Based on 1 review)

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 Human CXCL5/ENA-78 Quantikine ELISA Kit DX000 Array DX000
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 Human CXCL5/ENA-78 Quantikine ELISA Kit DX000
: Human CXCL5/ENA-78 Quantikine ELISA Kit [DX000].

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Sample Tested Cell Culture Media

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Other Experimental Details Curve fit in Prism and interpolated the sample concentrations. Kit worked well.

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