Human IL-8/CXCL8 QuantiGlo ELISA Kit

Catalog # Availability Size / Price Qty
Q8000B
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Human CXCL8/IL-8 Chemiluminescent ELISA Standard Curve
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Citations (10)
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Human IL-8/CXCL8 QuantiGlo ELISA Kit Summary

Assay Length
5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Citrate Plasma (50 uL)
Sensitivity
0.97 pg/mL
Assay Range
1.6 - 5,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Specificity
Natural and recombinant human IL-8
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 QuantiGlo Human IL-8 Chemiluminescent Immunoassay is a 5 hour solid phase ELISA designed to measure human IL-8 levels in cell culture supernates, serum, and plasma. It contains the 72 amino acid variant of human IL-8 derived from E. coli as well as antibodies raised against this recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human IL-8. Results obtained using natural human IL-8 showed linear curves that were parallel to the standard curves obtained using the QuantiGlo kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-8.

Precision

Intra-Assay Precision (Precision within an assay) Four samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Four samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 4 1 2 3 4
n 20 20 20 20 20 20 20 20
Mean 3.66 22.5 228 2263 4.19 24.9 243 2393
Standard Deviation 0.241 1.02 8.98 146 0.481 1.57 14.7 216
CV% 6.6 4.5 3.9 6.5 11.5 6.3 6 9

Recovery

The recovery of IL-8 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=4) 103 98-107
Citrate Plasma (n=4) 101 97-107
EDTA Plasma (n=4) 105 95-113
Heparin Plasma (n=4) 107 102-113
Serum (n=4) 103 93-107

Linearity

To assess the linearity of the assay, samples spiked with high concentrations of IL-8 in various matrices were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CXCL8/IL-8 Chemiluminescent ELISA Linearity

Data Examples

Human CXCL8/IL-8 Chemiluminescent ELISA Standard Curve

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: IL-8/CXCL8

Interleukin-8 (IL-8), also known as IL-8, GCP-1, and NAP-1, is a heparin-binding 8-9 kDa member of the alpha, or CXC family of chemokines. There are at least 15 human CXC family members that all adopt a three beta -sheet/one alpha -helix structure. Most CXC chemokines show an N-terminal Glu-Leu-Arg (ELR) tripeptide motif. IL-8 circulates as a monomer, homodimer, and heterodimer with CXCL4/PF4. The monomer is considered the most bio-active, while the heterodimer can potentiate PF4 activity. IL-8 oligomerization is modulated by its interactions with matrix and cell surface glycosaminoglycans (GAGs). Mature human IL-8 shares 65-69% amino acid (aa) identity with canine, feline, and porcine IL-8. There is no IL-8 gene counterpart in rodent. 


Multiple isoforms of IL-8 are generated through both alternative splicing and differential proteolytic cleavage. In humans, alternative splicing generates an iso-form with an eleven aa substitution at the C-terminus. Proteolytic processing results in N-terminal truncation of IL-8 and is likely a cell-specific event. For example, fibroblasts and endothelial cells generate the 1-77 form by cleaving IL-8 following Glu21, while monocytes and lymphocytes generate the 6-77 form by cleaving following Leu25. These truncated forms generally show increased bioactivity, particularly through the CXCR1 receptor. IL-8 can also undergo citrullination on Arg27 of the precursor, a modification that increases its half-life and ability to induce leukocytosis. A wide variety of cells secrete IL-8 including monocytes and neutrophils, fibroblasts and keratinocytes, mast cells, visceral smooth muscle cells, dendritic cells, type II great alveolar cells, and endothelial cells. 

IL-8 bioactivity is mediated through two G-protein-coupled receptors, termed CXCR1/IL-8 RA and CXCR2/IL-8 RB. CXCR1 is 45-50 kDa in size and is used almost exclusively by IL-8. CXCR2 is 35-40 kDa in size and is used by nearly all CXC chemokines. Both CXCR1 and CXCR2 constitutively associate into functional homodimers. They can also heterodimerize, but these complexes dissociate following IL-8 binding. CXCR2 responds to low concentrations of IL-8 and is principally associated with chemotaxis and MMP-9 release. CXCR1, in contrast, responds to high concentrations of IL-8 and is associated with respiratory burst and phospholipase D2 activation. Thus, CXCR2 ligation induces leukocyte adhesion to activated vascular endothelium and migration to sites of inflammation, while CXCR1 ligation primes neutrophil antimicrobial activity. IL-8 can also form a complex with Serpin A1/alpha-1 Antitrypsin, and this prevents IL-8 interaction with CXCR1. 

In addition to its pro-inflammatory effects, IL-8 is involved in angiogenesis and the pathogenesis of atherosclerosis and cancer. It induces VEGF expression in vascular endothelial cells and functions as an autocrine factor for EC growth and angiogenesis. It is upregulated in atherosclerotic lesions and is elevated in the serum and cerebrospinal fluid following myocardial infarction. In cancer, IL-8 promotes epithelial-mesenchymal transition as well as tumor cell invasiveness and metastasis.

Long Name:
Interleukin 8
Entrez Gene IDs:
3576 (Human); 396880 (Porcine); 403850 (Canine); 493836 (Feline)
Alternate Names:
3-10C; AMCF-I; C-X-C motif chemokine 8; CXCL8; CXCL8SCYB8; Emoctakin; GCP1; GCP-1TSG-1; IL8; IL-8; interleukin 8; K60; LAI; LECT; LUCT; LYNAP; MDNCF; MDNCFb-ENAP; member 8; MONAP; MONAPGCP1; NAF; NAP1; NAP-1NAP1; NCF; Neutrophil-activating protein 1; Protein 3-10C; T cell chemotactic factor; T-cell chemotactic factor; TCF; TSG1

Citations for Human IL-8/CXCL8 QuantiGlo 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.

10 Citations: Showing 1 - 10
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  1. MicroRNA-874 inhibits TNF-?-induced remodeling in human fetal airway smooth muscle cells by targeting STAT3
    Authors: M Sun, Y Huang, F Li, H Li, B Zhang, L Jin
    Respir Physiol Neurobiol, 2018;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Downregulation of tumor suppressive microRNAs in vivo in dense breast tissue of postmenopausal women
    Authors: A Abrahamsso, A Capodanno, A Rzepecka, C Dabrosin
    Oncotarget, 2017;8(54):92134-92142.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Activation of nuclear factor-kappa B signalling promotes cellular senescence.
    Authors: Rovillain E, Mansfield L, Caetano C, Alvarez-Fernandez M, Caballero OL, Medema RH, Hummerich H, Jat PS
    Oncogene, 2011;30(20):2356-66.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Procalcitonin as an early marker of bacterial infection in neutropenic febrile children with acute lymphoblastic leukemia.
    Authors: Hatzistilianou M, Rekliti A, Athanassiadou F, Catriu D
    Inflamm. Res., 2010;59(5):339-47.
    Species: Human
    Sample Types: Serum
  5. Phase I clinical and pharmacodynamic evaluation of oral CI-1033 in patients with refractory cancer.
    Authors: Zinner RG, Nemunaitis J, Eiseman I, Shin HJ, Olson SC, Christensen J, Huang X, Lenehan PF, Donato NJ, Shin DM
    Clin. Cancer Res., 2007;13(10):3006-14.
    Species: Human
    Sample Types: Plasma
  6. Presence of cytokeratins in exhaled breath condensate of mechanical ventilated patients.
    Authors: Gessner C, Dihazi H, Brettschneider S, Hammerschmidt S, Kuhn H, Eschrich K, Keller T, Engelmann L, Sack U, Wirtz H
    Respir Med, 2007;102(2):299-306.
    Species: Human
    Sample Types: Whole Cells
  7. Trichomonas vaginalis lipophosphoglycan triggers a selective upregulation of cytokines by human female reproductive tract epithelial cells.
    Authors: Fichorova RN, Trifonova RT, Gilbert RO, Costello CE, Hayes GR, Lucas JJ, Singh BN
    Infect. Immun., 2006;74(10):5773-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Recombinant human antithrombin inhibits thrombin formation and interleukin 6 release in human endotoxemia.
    Authors: Leitner JM, Firbas C, Mayr FB, Reiter RA, Steinlechner B, Jilma B
    Clin. Pharmacol. Ther., 2006;79(1):23-34.
    Species: Human
    Sample Types: Plasma
  9. Endurance training reduces circulating inflammatory markers in persons at risk of coronary events: impact on plaque stabilization?
    Authors: Niessner A, Richter B, Penka M, Steiner S, Strasser B, Ziegler S, Heeb-Elze E, Zorn G, Leitner-Heinschink A, Niessner C, Wojta J, Huber K
    Atherosclerosis, 2005;186(1):160-5.
    Species: Human
    Sample Types: Plasma
  10. Passive transfer of scrub typhus plasma to patients with AIDS: a descriptive clinical study.
    Authors: Watt G, Kantipong P, Jongsakul K, de Souza M, Burnouf T
    QJM, 2001;94(11):599-607.
    Species: Human
    Sample Types: Plasma

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