Human IL-6 ELISA - Quantikine QuicKit

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Human IL-6 ELISA Standard Curve
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Human IL-6 Quantikine QuicKit ELISA Summary

Assay Length
80 minutes
Sample Type & Volume Required Per Well
Cell Culture Supernates (10 uL), Serum (25 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL)
2.95 pg/mL
Assay Range
12.5 - 800 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Natural and recombinant human IL-6.
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.

Sample Values

Serum/Plasma - Ten serum and plasma samples from apparently healthy volunteers were evaluated for the presence of human IL-6 in this assay. All samples measured less than 25 pg/mL. No medical histories were available for the donors used in this study.

Cell Culture Supernates - Human peripheral blood mononuclear cells (PBMCs) (1 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were left unstimulated or stimulated with 1.0 μg/mL LPS for 24 hours. Aliquots of the culture supernates were removed, aassayed for levels of human IL-6, and were not detectable or measured 6356 pg/mL, respectively.

Monocytes were prepared from human PBMCs by adherence to plastic. Unattached cells were removed and attached monocytes were cultured in RPMI supplemented with 10% fetal bovine serum. Cells were left unstimulated or stimulated with 500 ng/mL LPS for 24 hours. Aliquots of the cell culture supernates were removed, assayed for levels of human IL-6, and were not detectable or measured 15,467 pg/mL, respectively.

Product Summary

The Quantikine® QuicKit™ Human IL-6 Immunoassay is a one step, 80-minute solid phase ELISA designed to measure human IL-6 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human IL-6 and antibodies raised against the recombinant protein. Results obtained using natural human IL-6 showed linear curves that were parallel to the standard curves obtained using the recombinant QuicKit™ standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-6.


The recovery of human IL-6 spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated. 

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 119 114-126
EDTA Plasma (n=2) 117 112-122
Heparin Plasma (n=2) 111 107-114
Serum (n=2) 106 96-115


To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human IL-6 in various matrices and diluted to produce samples with values within the dynamic range of the assay. 
Human IL-6 ELISA Linearity

Scientific Data

Human IL-6 ELISA Standard Curve

Human IL-6 QuicKit Spiked Recovery Competitor Comparison IL-6 is spiked at three known concentrations throughout the range of the assay and run to measure response of the spiked sample matrix. Serum recovery is 109% compared to 68% for the top competitor. Plasma recovery is 113% compared to 77% for the top competitor.

Human IL-6 QuicKit Spiked Linearity Competitor Comparison IL-6 is spiked at high concentration in various matrices and diluted with appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay. The linearity is between 86%-107% compared to 111%-149% for the top competitor.

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IL-6

Interleukin 6 (IL-6) is a pleiotropic, a-helical, 22-28 kDa phosphorylated and variably glycosylated cytokine that plays important roles in the acute phase reaction, inflammation, hematopoiesis, bone metabolism, and cancer progression. Mature human IL-6 is 183 amino acids (aa) in length and shares 39% aa sequence identity with mouse and rat IL-6. Alternative splicing generates several isoforms with internal deletions, some of which exhibit antagonistic properties. Cells known to express IL-6 include CD8+ T cells, fibroblasts, synoviocytes, adipocytes, osteoblasts, megakaryocytes, endothelial cells (under the influence of endothelins), sympathetic neurons, cerebral cortex neurons, adrenal medulla chromaffin cells, retinal pigment cells, mast cells, keratinocytes, Langerhans cells, fetal and adult astrocytes, neutrophils, monocytes, eosinophils, colonic epithelial cells, B1 B cells and pancreatic islet beta cells. IL-6 production is generally correlated with cell activation and is normally kept in control by glucocorticoids, catecholamines, and secondary sex steroids. Normal human circulating IL-6 is in the 1 pg/mL range, with slight elevations during the menstrual cycle, modest elevations in certain cancers, and large elevations after surgery.

IL-6 induces signaling through a cell surface heterodimeric receptor complex composed of a ligand binding subunit (IL-6 R alpha) and a signal transducing subunit (gp130). IL-6 binds to IL-6 Ra, triggering IL-6 Ra association with gp130 and gp130 dimerization. gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM. Soluble forms of IL-6 Ra are generated by both alternative splicing and proteolytic cleavage. In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 Ra elicit responses from gp130-expressing cells that lack cell surface IL-6 Ra. Trans-signaling enables a wider range of cell types to respond to IL-6, as the expression of gp130 is ubiquitous, while that of IL-6 Ra is predominantly restricted to hepatocytes, monocytes, and resting lymphocytes. Soluble splice forms of gp130 block trans-signaling from IL-6/IL-6 Ra but not from other cytokines that use gp130 as a co-receptor. 

IL-6, along with TNF-a and IL-1, drives the acute inflammatory response. IL-6 is almost solely responsible for fever and the acute phase response in the liver, and it is important in the transition from acute inflammation to either acquired immunity or chronic inflammatory disease. When dysregulated, it contributes to chronic inflammation in conditions such as obesity, insulin resistance, inflammatory bowel disease, arthritis, and sepsis. IL-6 modulates bone resorption and is a major effector of inflammatory joint destruction in rheumatoid arthritis through its promotion of Th17 cell development and activity. It contributes to atherosclerotic plaque development and destabilization as well as the development of inflammation-associated carcinogenesis.

Long Name:
Interleukin 6
Entrez Gene IDs:
3569 (Human); 16193 (Mouse); 24498 (Rat); 399500 (Porcine); 280826 (Bovine); 403985 (Canine); 102138971 (Cynomolgus Monkey); 100034196 (Equine); 493687 (Feline); 463288 (Primate); 100008733 (Rabbit)
Alternate Names:
B-cell differentiation factor; B-cell stimulatory factor 2; BSF2; BSF-2; CDF; CTL differentiation factor ; HSF; hybridoma growth factor; IFNB2; IFN-beta-2; IL6; IL-6; Interferon beta-2; interleukin 6 (interferon, beta 2); interleukin BSF-2; interleukin-6; MGI-2A

Assay Procedure

These assays utilize an anti-tag coated microplate. Standards, samples, and controls are added to plate, followed by subsequent addition of an antibody cocktail. Following a 1 hour incubation, plates are washed and substrate is added and incubated for 20 minutes. Stop solution is then added and plates are read using a standard microplate reader.

QuicKit ELISA assay procedure

Citations for Human IL-6 Quantikine QuicKit ELISA

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.

3 Citations: Showing 1 - 3
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  1. Unravelling heterogeneous effects of cancer?associated fibroblasts on poor prognosis markers in breast cancer EM?G3 cell line: In vitro?targeted treatment (anti?IL-6, anti?VEGF-A, anti?MFGE8) based on transcriptomic profiling
    Authors: Urban, L;Novák, Š;?oma, M;Dvo?ánková, B;Lacina, L;Šáchová, J;Hradilová, M;Svato?ová, P;Kolá?, M;Strnad, H;B?ezinová, J;Smetana, K;Gál, P;Szabo, P;
    Oncology reports
    Species: Human
    Sample Types: Cell Culture Supernates
  2. LINC01094/SPI1/CCL7 Axis Promotes Macrophage Accumulation in Lung Adenocarcinoma and Tumor Cell Dissemination
    Authors: Z Wu, X Bai, Z Lu, S Liu, H Jiang
    Journal of Immunology Research, 2022-09-09;2022(0):6450721.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. 4-octyl Itaconate�inhibits lipopolysaccharide (LPS)-induced osteoarthritis via activating Nrf2 signalling pathway
    Authors: Q Zhang, X Bai, R Wang, H Zhao, L Wang, J Liu, M Li, Z Chen, Z Wang, L Li, D Wang
    Journal of Cellular and Molecular Medicine, 2022-01-23;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates


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Human IL-6 Quantikine QuicKit ELISA
By James AHODANTIN on 01/08/2022
Sample Tested: Human cell conditioned medium

Very good and sensitive kit. I used 1/1000 diluted conditioned media from macrophages stimulated with LPS. Very reliable kit.