Human IL-8/CXCL8 Quantikine HS ELISA Kit

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HS800
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Human CXCL8/IL-8 ELISA Standard Curve
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Human IL-8/CXCL8 Quantikine HS ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
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 Quantikine HS Human IL-8 Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-8 levels in serum and plasma. It contains E. coli-expressed recombinant human IL-8 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant human IL-8 accurately. Results obtained using natural human IL-8 showed linear curves that were parallel to the standard curves obtained using the Quantikine HS 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) 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

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 5.5 18.7 37.1 5 19.4 39.2
Standard Deviation 0.3 0.7 2.7 0.4 1.6 3.7
CV% 5.5 3.7 7.3 8 8.2 9.4

Recovery

The recovery of IL-8 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
EDTA Plasma (n=4) 95 86-102
Heparin Plasma (n=4) 99 91-107
Serum (n=4) 99 85-110

Linearity

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

Data Examples

Human CXCL8/IL-8 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

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. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 100 µL Standard, Control, or Sample
  6.   Add 100 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 5 times for a total of 6 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 1 hour.
  10.   Aspirate and wash 6 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour. Do not wash the plate.

  13. 200 µL Amplifier Solution
  14.   Add 200 µL Amplifier Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.

  15. 50 µL Stop Solution
  16. Add 50 µL of Stop Solution to each well. Read at 490 nm within 30 minutes. Set wavelength correction to 650 nm or 690 nm.

Citations for Human IL-8/CXCL8 Quantikine HS 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.

17 Citations: Showing 1 - 10
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  1. The relationship between malnutrition risk and inflammatory biomarkers in outpatient geriatric population
    Authors: P Fatyga, A Pac, M Fedyk-?uka, T Grodzicki, A Skalska
    Eur Geriatr Med, 2020;0(0):.
    Species: Human
    Sample Types: Serum
  2. HIV antiretroviral drugs, dolutegravir, maraviroc and ritonavir-boosted atazanavir use different pathways to affect inflammation, senescence and insulin sensitivity in human coronary endothelial cells
    Authors: M Auclair, AC Guénantin, S Fellahi, M Garcia, J Capeau
    PLoS ONE, 2020;15(1):e0226924.
    Species: Human
    Sample Types: cell culture supernatant
  3. LIPUS vs. reaming in non-union treatment: Cytokine expression course as a tool for evaluation and differentiation of non-union therapy
    Authors: J Doll, A Moghaddam, V Daniel, B Biglari, R Heller, G Schmidmaie, TF Raven
    J Orthop, 2020;17(0):208-214.
    Species: Human
    Sample Types: Serum
  4. Biological variation of immunological blood biomarkers in healthy individuals and quality goals for biomarker tests
    Authors: N Aziz, R Detels, JJ Quint, D Gjertson, T Ryner, AW Butch
    BMC Immunol., 2019;20(1):33.
    Species: Human
    Sample Types: Serum
  5. Interleukin-8 as a therapeutic target for chronic low back pain: Upregulation in human cerebrospinal fluid and pre-clinical validation with chronic reparixin in the SPARC-null mouse model
    Authors: E Krock, M Millecamps, KM Anderson, A Srivastava, TE Reihsen, P Hari, YR Sun, SH Jang, GL Wilcox, KG Belani, DS Beebe, J Ouellet, MR Pinto, LJ Kehl, L Haglund, LS Stone
    EBioMedicine, 2019;0(0):.
    Species: Human
    Sample Types: CSF
  6. Impact of constitutional TET2 haploinsufficiency on molecular and clinical phenotype in humans
    Authors: E Kaasinen, O Kuismin, K Rajamäki, H Ristolaine, M Aavikko, J Kondelin, S Saarinen, DG Berta, R Katainen, EAM Hirvonen, A Karhu, A Taira, T Tanskanen, A Alkodsi, M Taipale, E Morgunova, K Franssila, R Lehtonen, M Mäkinen, K Aittomäki, A Palotie, MI Kurki, O Pietiläine, M Hilpert, E Saarentaus, J Niinimäki, J Junttila, K Kaikkonen, P Vahteristo, RC Skoda, MRJ Seppänen, KK Eklund, J Taipale, O Kilpivaara, LA Aaltonen
    Nat Commun, 2019;10(1):1252.
    Species: Human
    Sample Types: Plasma
  7. Sex-specific effects of central adiposity and inflammatory markers on limbic microstructure
    Authors: C Metzler-Ba, JP Mole, E Leonaviciu, R Sims, EJ Kidd, B Ertefai, A Kelso-Mitc, F Gidney, F Fasano, J Evans, DK Jones, RJ Baddeley
    Neuroimage, 2019;189(0):793-803.
    Species: Human
    Sample Types: Plasma
  8. The influence of prostatic Cutibacterium acnes infection on serum levels of IL6 and CXCL8 in prostate cancer patients
    Authors: H Ugge, J Carlsson, B Söderquist, K Fall, O Andén, S Davidsson
    Infect. Agents Cancer, 2018;13(0):34.
    Species: Human
    Sample Types: Serum
  9. Pilot study of placental tissue collection, processing, and measurement procedures for large scale assessment of placental inflammation
    Authors: LA Sjaarda, KA Ahrens, DL Kuhr, TL Holland, UR Omosigho, BT Steffen, NL Weir, HK Tollman, RM Silver, MY Tsai, EF Schisterma
    PLoS ONE, 2018;13(5):e0197039.
    Species: Human
    Sample Types: Tissue Homogenates
  10. Mindfulness and its efficacy for psychological and biological responses in women with breast cancer
    Authors: E Kenne Sare, LB Mårtensson, BA Andersson, P Karlsson, I Bergh
    Cancer Med, 2017;0(0):.
    Species: Human
    Sample Types: Serum
  11. Microenvironment inflammatory infiltrate drives growth speed and outcome of hepatocellular carcinoma: a prospective clinical study
    Authors: R Critelli, F Milosa, F Faillaci, R Condello, E Turola, L Marzi, B Lei, F Dituri, S Andreani, P Sighinolfi, P Manni, A Maiorana, C Caporali, F di Benedet, M Del Buono, N De Maria, F Schepis, ML Martinez-C, G Giannelli, E Villa
    Cell Death Dis, 2017;8(8):e3017.
    Species: Human
    Sample Types: Serum
  12. Exogenous growth factors bFGF, EGF and HGF do not influence viability and phenotype of V600EBRAF melanoma cells and their response to vemurafenib and trametinib in vitro
    Authors: I Zalesna, M Osrodek, ML Hartman, M Rozanski, M Sztiller-S, K Niewinna, D Nejc, M Czyz
    PLoS ONE, 2017;12(8):e0183498.
    Species: Human
    Sample Types: Whole Cells
  13. Circulating mitochondrial DNA increases with age and is a familiar trait: Implications for "inflamm-aging"
    Authors: Pinti M, Cevenini E, Nasi M, De Biasi S, Salvioli S, Monti D, Benatti S, Gibellini L, Cotichini R, Stazi M, Trenti T, Franceschi C, Cossarizza A
    Eur J Immunol, 2014;44(5):1552-62.
    Species: Human
    Sample Types: Plasma
  14. Inflammatory thresholds and the species-specific effects of colonising bacteria in stable chronic obstructive pulmonary disease.
    Authors: Singh R, Mackay A, Patel A, Garcha D, Kowlessar B, Brill S, Donnelly L, Barnes P, Donaldson G, Wedzicha J
    Respir Res, 2014;15(0):114.
    Species: Human
    Sample Types: Sputum
  15. The expression of cytokines and chemokines in the blood of patients with severe weight loss from anorexia nervosa: an exploratory study.
    Authors: Pisetsky D, Trace S, Brownley K, Hamer R, Zucker N, Roux-Lombard P, Dayer J, Bulik C
    Cytokine, 2014;69(1):110-5.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Endothelial activation by platelets from sickle cell anemia patients.
    Authors: Proenca-Ferreira R, Brugnerotto A, Garrido V, Dominical V, Vital D, Ribeiro M, dos Santos M, Traina F, Olalla-Saad S, Costa F, Conran N
    PLoS ONE, 2014;9(2):e89012.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Association between traffic-related air pollution, subclinical inflammation and impaired glucose metabolism: results from the SALIA study.
    Authors: Teichert T, Vossoughi M, Vierkotter A, Sugiri D, Schikowski T, Schulte T, Roden M, Luckhaus C, Herder C, Kramer U
    PLoS ONE, 2013;8(12):e83042.
    Species: Human
    Sample Types: Plasma

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