Summary
Precision
Recovery
Linearity
Data Examples
Product Datasheets
Preparation and Storage
Background
Related Research Areas
Human CXCL6/GCP-2 Quantikine ELISA Kit Summary
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 (25 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Sensitivity
8 pg/mL
Assay Range
31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human GCP-2
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 GCP-2 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human GCP-2 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human GCP-2 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human GCP-2 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 GCP-2.
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, Serum, EDTA Plasma, Heparin Plasma
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 273 | 517 | 1078 | 263 | 472 | 965 |
| Standard Deviation | 13.7 | 30.5 | 57.7 | 20.7 | 35.5 | 65.6 |
| CV% | 5 | 5.9 | 5.4 | 7.9 | 7.5 | 6.8 |
Recovery
The recovery of GCP-2 spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 104 | 93-114 |
| EDTA Plasma (n=5) | 99 | 89-109 |
| Heparin Plasma (n=5) | 100 | 91-112 |
| Serum (n=5) | 95 | 88-104 |
Linearity
To assess the linearity of the assay, samples containing or spiked with high concentrations of GCP-2 were serially diluted with the Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data
Product Datasheets
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CXCL6/GCP-2
GCP-2 (granulocyte chemotactic protein-2) is a CXC chemokine. Among human CXC chemokines, GCP-2 is most closely related to ENA-78. The structure and sequence of the genes for human GCP-2 and ENA-78 also exhibit close similarity, suggesting the two genes may have originated from a recent gene duplication. LIX (LPS-induced CXC chemokine) was initially cloned as a gene induced by LPS in mouse fibroblasts. The mouse protein designated GCP-2, because of its amino acid sequence similarity (60%) to human GCP-2, is identical to the LIX protein sequence.
Entrez Gene IDs:
6372 (Human)
Alternate Names:
chemokine (C-X-C motif) ligand 6 (granulocyte chemotactic protein 2); Chemokine alpha 3; CKA-3Granulocyte chemotactic protein 2; CXCL6; GCP-2; GCP2member 6 (granulocytechemotactic protein 2); GCP-2Small-inducible cytokine B6; Small inducible cytokine subfamily B (Cys-X-Cys), member b
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.
Assay Procedure
Refer to the product Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- 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.
100 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Citations for Human CXCL6/GCP-2 Quantikine ELISA Kit
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 - 8
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Human cardiac progenitor cell activation and regeneration mechanisms: exploring a novel myocardial ischemia/reperfusion in vitro model
Authors: MJ Sebastião, M Serra, R Pereira, I Palacios, P Gomes-Alve, PM Alves
Stem Cell Res Ther, 2019-03-07;10(1):77.
Species: Human
Sample Types: Cell Culture Supernates
-
Ets Homologous Factor (EHF) has Critical Roles in Epithelial Dysfunction in Airway Disease
Authors: SL Fossum, MJ Mutolo, A Tugores, S Ghosh, SH Randell, LC Jones, SH Leir, A Harris
J. Biol. Chem., 2017-05-01;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2015-04-28;10(4):e0124283.
Species: Human
Sample Types: Cell Culture Supernates
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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., 2007-08-20;38(2):168-75.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2006-10-04;192(1):56-66.
Species: Human
Sample Types: Cell Culture Supernates
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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., 2005-11-01;175(9):6177-89.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2004-12-01;22(6):513-20.
Species: Human
Sample Types: Plasma
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ELR+ CXC chemokines in human milk.
Authors: Maheshwari A, Christensen RD, Calhoun DA
Cytokine, 2003-11-07;24(3):91-102.
Species: Human
Sample Types: Serum
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