The Human IL-6 ELISA kit is a single-wash, 90-minute SimpleStep ELISA® assay designed for the quantification of human interleukin-6 (IL-6) with a sensitivity of 1.6 pg/ml. The assay uses a simple Mix-Wash-Read protocol with just one incubation and wash step.Easy-to-use, rapid, sensitive human IL-6 sandwich ELISA kit:- Colorimetric assay - 450nm readout; works on any plate reader- Different formats for different needs: 10x96 plates for bulk orders and 384-well for higher throughput- Versatile sample use – Compatible with serum, plasma, and cell culture supernatants
Colorimetric
Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma, Cerebral Spinal Fluid
Sandwich (quantitative)
Human
7.8 - 500 pg/mL
1h 30m
= 1.6 pg/mL
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Cytokine with a wide variety of biological functions in immunity, tissue regeneration, and metabolism. Binds to IL6R, then the complex associates to the signaling subunit IL6ST/gp130 to trigger the intracellular IL6-signaling pathway (Probable). The interaction with the membrane-bound IL6R and IL6ST stimulates 'classic signaling', whereas the binding of IL6 and soluble IL6R to IL6ST stimulates 'trans-signaling'. Alternatively, 'cluster signaling' occurs when membrane-bound IL6:IL6R complexes on transmitter cells activate IL6ST receptors on neighboring receiver cells (Probable).IL6 is a potent inducer of the acute phase response. Rapid production of IL6 contributes to host defense during infection and tissue injury, but excessive IL6 synthesis is involved in disease pathology. In the innate immune response, is synthesized by myeloid cells, such as macrophages and dendritic cells, upon recognition of pathogens through toll-like receptors (TLRs) at the site of infection or tissue injury (Probable). In the adaptive immune response, is required for the differentiation of B cells into immunoglobulin-secreting cells. Plays a major role in the differentiation of CD4(+) T cell subsets. Essential factor for the development of T follicular helper (Tfh) cells that are required for the induction of germinal-center formation. Required to drive naive CD4(+) T cells to the Th17 lineage. Also required for proliferation of myeloma cells and the survival of plasmablast cells (By similarity).Acts as an essential factor in bone homeostasis and on vessels directly or indirectly by induction of VEGF, resulting in increased angiogenesis activity and vascular permeability (PubMed:12794819, PubMed:17075861). Induces, through 'trans-signaling' and synergistically with IL1B and TNF, the production of VEGF (PubMed:12794819). Involved in metabolic controls, is discharged into the bloodstream after muscle contraction increasing lipolysis and improving insulin resistance (PubMed:20823453). 'Trans-signaling' in central nervous system also regulates energy and glucose homeostasis (By similarity). Mediates, through GLP-1, crosstalk between insulin-sensitive tissues, intestinal L cells and pancreatic islets to adapt to changes in insulin demand (By similarity). Also acts as a myokine (Probable). Plays a protective role during liver injury, being required for maintenance of tissue regeneration (By similarity). Also has a pivotal role in iron metabolism by regulating HAMP/hepcidin expression upon inflammation or bacterial infection (PubMed:15124018). Through activation of IL6ST-YAP-NOTCH pathway, induces inflammation-induced epithelial regeneration (By similarity).
IFNB2, IL6, Interleukin-6, IL-6, B-cell stimulatory factor 2, CTL differentiation factor, Hybridoma growth factor, Interferon beta-2, BSF-2, CDF, IFN-beta-2
The Human IL-6 ELISA kit is a single-wash, 90-minute SimpleStep ELISA® assay designed for the quantification of human interleukin-6 (IL-6) with a sensitivity of 1.6 pg/ml. The assay uses a simple Mix-Wash-Read protocol with just one incubation and wash step.Easy-to-use, rapid, sensitive human IL-6 sandwich ELISA kit:- Colorimetric assay - 450nm readout; works on any plate reader- Different formats for different needs: 10x96 plates for bulk orders and 384-well for higher throughput- Versatile sample use – Compatible with serum, plasma, and cell culture supernatants
Colorimetric
Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma, Cerebral Spinal Fluid
Sandwich (quantitative)
Human
7.8 - 500 pg/mL
1h 30m
Microplate (12 x 8 well strips)
= 1.6 pg/mL
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Supernatant | n 5 | mean - | SD - | C.V. 2.1 |
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Supernatant | n 3 | mean - | SD - | C.V. 2.4 |
Sample type | Average % | Range |
---|---|---|
Sample type Cell culture media | Average % = 100 | Range 97 - 103 % |
Sample type Cerebral Spinal Fluid | Average % = 90 | Range 87 - 93 % |
Sample type Citrate plasma | Average % = 84 | Range 81 - 85 % |
Sample type EDTA Plasma | Average % = 82 | Range 80 - 85 % |
Sample type Serum | Average % = 82 | Range 77 - 84 % |
Blue Ice
+4°C
+4°C
+4°C
Human IL-6 ELISA kit ab178013 is a rapid single-wash 90-min ELISA kit to measure Human IL-6 in cerebral spinal fluid (CSF), cell culture supernatant, serum and plasma. This SimpleStep ELISA® sensitivity is 1.6 pg/mL.
How the assay works
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details.
Assay protocol summary
Human IL-6 ELISA kit ab178013 protocol summary:
1. Mix : add samples / standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hour at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm
Alternative assay kits
Need to design your own immunoassay? We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:
- Human/Monkey IL-6 Antibody Pair - BSA and Azide free Human/Monkey IL-6 Antibody Pair - BSA and Azide free ab243973,
- Anti-IL-6 antibody [IL-6-10] - BSA and Azide free (Capture) Anti-IL-6 antibody [IL-6-10] - BSA and Azide free (Capture) ab244758,
- Anti-IL-6 antibody [IL-6-5] - BSA and Azide free (Detector) Anti-IL-6 antibody [IL-6-5] - BSA and Azide free (Detector) ab244886
Getting the best performance
ASSAY SPECIFICITY
This kit recognizes both native and recombinant Human IL-6 protein in serum, plasma, and cell culture supernatant samples only. Cell and tissue extract samples have not been tested with this kit.
CROSS REACTIVITY
Recombinant Human IL-6Ra was prepared at 50 ng/mL and 500 pg/mL and assayed for cross reactivity. No cross-reactivity was observed.
INTERFERENCE
Recombinant Human IL-6Ra was prepared at 50 ng/mL and 500 pg/mL and tested for interference. No interference with was observed.
SPECIES REACTIVITY
This kit recognizes Human IL-6 protein. Recombinant mouse IL-6 was prepared at 500 pg/mL and assayed for cross reactivity. No cross-reactivity was observed. Please contact our Technical Support team for more information.
Save your precious samples
Save your precious samples: our 384-well Human IL-6 ELISA kit only requires a maximum of 12.5µL
How other researchers are using this kit
Human IL-6 ELISA kit ab178013 has been used to assess inflammation in different physio-pathological contexts such as obesity1, coronary artery disease2, and rheumatoid arthritis3.
References: 1-Shaohong Yu et al. 2023, PMID: 38156278, 2-Yongshun W. et al. 2023, PMID:38054817, 3-Yaju Tan et al. 2024, PMID: 37997635
Related and recommended products
Human IL-6 ELISA kit ab178013 is commonly used to measure IL-6 as a marker of inflammation.
Other ELISA kits to measure inflammatory markers include:
-Human TNF alpha ELISA kit Human TNF alpha ELISA Kit ab181421
- Human IL-8 ELISA kit Human IL-8 ELISA Kit ab214030
- Human IL-1 beta ELISA kit Human IL-1 beta ELISA Kit ab214025
In glioblastoma, IL-6 is known to activate several pro-oncogenic signaling pathways.
Explore our comprehensive range of glioblastoma research solutions here: https://www.abcam.com/en-us/products/product-recommendations/glioblastoma-research-tools
In Alzheimer's disease, increased levels of IL-6 in the plasma or cerebrospinalfluid ofpatients is associated with cognitive impairement.
Explore our extensive Alzheimer's research solutions here: https://www.abcam.com/en-us/products/product-recommendations/alzheimers-disease-tools
This supplementary information is collated from multiple sources and compiled automatically.
Interleukin-6 (IL-6) also known as B-cell stimulatory factor-2 or interferon beta-2 is a multifunctional cytokine with a molecular weight of approximately 26 kDa. It is expressed in a variety of tissues including T cells macrophages and vascular endothelial cells. IL-6 has a role in immune response and inflammation by mediating the actions between lymphocytes and non-lymphoid cells. Researchers can measure IL-6 levels using specialized assays like the IL-6 ELISA or through employing IL-6 recombinant protein.
IL-6 is significant in immune regulation and inflammation. IL-6 which often functions as a single molecule or in complex with the soluble IL-6 receptor mediates its effects through binding to the IL-6 receptor complex and gp130 protein which triggers intracellular signaling cascades. This protein contributes significantly to processes like B cell differentiation into plasma cells as well as liver regeneration and metabolic regulation.
IL-6 actively participates in critical signaling pathways such as the JAK/STAT pathway and the MAPK pathway. Upon receptor binding it engages the JAK kinases which then phosphorylate and activate STAT proteins leading to gene expression changes. The MAPK pathway activation involves proteins like Ras and Raf ultimately leading to downstream effects like cell growth and differentiation. Together these pathways illustrate the versatile roles of IL-6 in immune responses and inflammation.
IL-6 connects particularly with inflammatory and autoimmune conditions. Elevated IL-6 levels associate with rheumatoid arthritis and chronic inflammatory conditions where it contributes to tissue inflammation and joint damage through mediators like TNF-alpha. In some cancers IL-6 promotes tumor growth and survival by stimulating angiogenesis. The link with TNF-alpha and other inflammatory cytokines positions IL-6 as an important target for therapeutic interventions in such disorders.
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Human IL6 standard curve comparison data.
Standard curve comparison between the previous Human IL6 SimpleStep ELISA® kit and the re-developed Human IL6 SimpleStep ELISA® kit. The re-developed kit shows increased sensitivity.
Example of human IL6 standard curve.
Background-subtracted data values (mean +/- SD) are graphed.
Example of raw data value for of human IL-6 standard curve.
Raw data values are shown in the table.
Interpolated concentrations of spiked IL6 in human serum and plasma samples.
The concentrations of IL6 were measured in duplicates, interpolated from the IL6 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (EDTA) 50%, and plasma (citrate) 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
Interpolated concentrations of native IL6 in human peripheral blood monocyte (PBMC) cell culture supernatant samples.
The concentrations of IL6 were measured in duplicates, interpolated from the IL6 standard curves and corrected for sample dilution. Undiluted samples are as follows: Stimulated PBMC Cell culture supernatant 1:200. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL6 concentration was determined to be 60,443 pg/mL in stimulated PBMC cell culture supernatant, and undetectable in unstimulated and media controls.
Linearity of dilution for spiked recombinant IL-6.
Recombinant IL-6 was spiked into biological samples and diluted in a 2-fold dilution series in Sample Diluent NS. NL= Non linear 50% pooled serum and plasma (EDTA, Citrate) samples from healthy donors was measured in duplicate. All values were below the detectable range of the assay.
50% serum from ten individual healthy human male and female donors was measured in duplicate. All values were below the detectable range of the assay.
Linearity of dilution for native IL-6.
Native IL-6 was measured in biological samples in a 2- fold dilution series. Sample dilutions are made in Sample Diluent NS.
Human IL-6 concentration was interpolated from the IL-6 standard curve. Supernatants from cell culture samples were serially diluted and assessed by the Human IL-6 ELISA kit (ab178013). Wild-type and IL-6 knockout A549 cells (Human IL-6 knockout A549 cell line ab273751) were assessed in duplicate (n=2). Cells were either treated with 20 ng/mL active recombinant human IL-1 beta protein (Recombinant human IL-1 beta protein (Active) ab259387) for 24 h to induce expression of IL-6 or not treated with IL-1 beta. Data are represented as the mean and error bars represent standard deviation.
Interpolated concentrations of Human IL-6 in human stimulated PBMC supernatant in 96-well and 384-well plates.
Interpolated concentration of native IL-6 was measured in duplicate at different sample concentrations in 96-well vs. 384-well plates. Undiluted samples are 2% Human Stimulated PBMC supernatant treated with 1.5% PHA-M for 46 hrs. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.
Example of Human IL-6 standard curve in Sample Diluent NS in 96-well and 384-well plate.
Example of Human IL-6 standard curve in 96-well and 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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