Human IL-8/CXCL8 ELISA Kit - Quantikine
R&D Systems, part of Bio-Techne | Catalog # D8000C
Key Product Details
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Citrate Plasma (50 µL)
Sensitivity
7.5 pg/mL
Assay Range
31.2-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Product Summary for Human IL-8/CXCL8 Quantikine ELISA Kit
The Quantikine Human IL-8 Immunoassay is a 3.5 hour solid phase ELISA designed to measure human IL-8 in cell culture supernates, serum, and plasma. It is based on antibodies raised against the 72 aa variant of human IL-8 derived from E. coli. It is calibrated with the same recombinant factor. This immunoassay accurately quantitates recombinant human IL-8. Measurement of natural human IL-8 or the 77 aa variant of human IL-8 gave results parallel to the standard curves obtained using the E. coli-expressed Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-8.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human
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.
Sample Values
Serum/Plasma - Thirty-four samples from apparently healthy volunteers were evaluated for
the presence of human IL-8 in this assay. All samples measured less than the lowest human IL-8
standard, 31.3 pg/mL. No medical histories were available for the donors used in this study.
Cell Culture Supernates - Human peripheral blood mononuclear cells (1 x 106
cells/mL) were
cultured in RPMI supplemented with 10% fetal bovine serum, 50 μM b-mercaptoethanol,
2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate and stimulated
with 10 μg/mL PHA. Aliquots of the culture supernate were removed on days 1 and 5 and
assayed for levels of human IL-8.
| Condition | Day 1 (pg/mL) | Day 5 (pg/mL) |
| Unstimulated | 27,000 | 33.000 |
| Stimulated | 73,000 | 102,000 |
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) hree 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 | 20 | 20 | 20 |
| Mean (pg/mL) | 115 | 386 | 802 | 132 | 410 | 817 |
| Standard Deviation | 5.3 | 17.0 | 37.9 | 10.7 | 28.0 | 42.4 |
| CV% | 4.6 | 4.4 | 4.7 | 8.1 | 6.8 | 5.2 |
Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 168 | 526 | 1093 | 196 | 581 | 1160 |
| Standard Deviation | 9.4 | 28.6 | 70.7 | 14.5 | 56.6 | 70.2 |
| CV% | 5.6 | 5.4 | 6.5 | 7.4 | 9.7 | 6.1 |
Recovery for Human IL-8/CXCL8 Quantikine ELISA Kit
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=5) | 98 | 85-114 |
| Citrate Plasma (n=5) | 105 | 95-114 |
| EDTA Plasma (n=5) | 103 | 97-111 |
| Heparin Plasma (n=5) | 102 | 92-107 |
| Serum (n=5) | 98 | 88-106 |
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of IL-8 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human IL-8/CXCL8 Quantikine ELISA Kit
Human CXCL8 ELISA/IL-8 ELISA Cell Culture Supernate Standard Curve
Human CXCL8 ELISA/IL-8 ELISA Serum/Plasma Standard Curve
Human IL-8/CXCL8 Quantikine ELISA Kit
RT-qPCR for a set of cytokines (CSF2, IL6, IL8 and IL1B) and chemokines (CXCL1 and CXCL6) of total RNA samples derived from mono- and co-cultures in a transwell assay.The respective co- (+) or mono- (-) culture is indicated on the X-axis. The respective analyzed cytokine or chemokine is indicated in the header of each graph. Expression values are shown in arbitrary units (AU) and have been normalized to beta-2 microglobulin (B2M) mRNA copies. Experiments were performed in triplicates (n = 3). Statistical analysis was performed on the mean values by unpaired comparison of mono-cultured HDF and co-cultured HDF RNA samples by using Student’s t-test (**p<0.01, ***p<0.001; n.s.: not significant). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25919140), licensed under a CC-BY license. Not internally tested by R&D Systems.
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-8/CXCL8
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
Alternate Names
CXCL8, GCP1, IL8, LAI, LECT, LUCT, LYNAP, MDNCF, MONAP, NAF, NAP1, NCF, TCF, TSG1
Gene Symbol
CXCL8
Additional IL-8/CXCL8 Products
Product Documents for Human IL-8/CXCL8 Quantikine ELISA Kit
Product Specific Notices for Human IL-8/CXCL8 Quantikine ELISA Kit
For research use only
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⚠ 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.