Mouse IL-6 Quantikine ELISA Kit

R&D Systems | Catalog # M6000B

2 plate pack size in process of discontinuation
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), Heparin Plasma (50 µL)

Sensitivity

1.8 pg/mL

Assay Range

7.8-500 pg/mL (Cell Culture Supernates, Serum, Heparin Plasma)
View all IL-6 ELISA Kits »

Product Summary for Mouse IL-6 Quantikine ELISA Kit

The Quantikine Mouse IL-6 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse IL-6 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse IL-6 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse IL-6. Results obtained using natural mouse IL-6 showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse IL-6.

Product Specifications

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Mouse

Specificity

Natural and recombinant mouse IL-6

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 - Twenty individual mouse serum samples were evaluated for the presence of mouse IL-6 in this assay. In all samples, IL-6 levels measured below the low standard, 7.8 pg/mL.

A mouse was injected with 15 μg of LPS and was bled 2 hours after injection. The serum was tested for mouse IL-6 and measured 31,800 pg/mL.
Heparin Plasma - Eight individual mouse heparin plasma samples were evaluated for the presence of mouse IL-6 in this assay. Six samples were below the low standard, 7.8 pg/mL. Two samples measured 23 pg/mL and 18 pg/mL.

Cell Culture Supernates: 
EL-4 mouse lymphoblast cells (1 x 106 cells/mL) were cultured for 3 days in DMEM supplemented with 10% fetal bovine serum and stimulated with 10 μg/mL PHA and 10 μg/mL PMA. The culture supernate was assayed for mouse IL-6 and measured 386 pg/mL.

D10.G4.1 mouse helper T cells (1 x 105 cells/mL) were cultured in RPMI 1640 supplemented with L-glutamine, 10% fetal bovine serum, 50 μM beta -mercaptoethanol, and 2.5% rat T cell polyclone media (Collaborative # 40115). During the final 3 days of culture, cells were stimulated with 2.5 ng/mL LPS. The culture supernate was assayed for mouse IL-6 and measured 759 pg/mL.

Mouse heart cells were cultured for 7 days in RPMI supplemented with 10% fetal bovine serum. The culture supernate was assayed for mouse IL-6 and measured 1120 pg/mL.



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.

Cell Culture Supernates, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 28 22 27
Mean (pg/mL) 30 87 232 29 85 238
Standard Deviation 2.0 3.4 8.1 1.8 7.5 18.1
CV% 6.7 3.9 3.5 6.1 8.9 7.6

Recovery for Mouse IL-6 Quantikine ELISA Kit

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

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=5) 110 86-120
Heparin Plasma (n=6) 101 81-115
Serum (n=5) 99 84-113

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse IL-6 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Mouse IL-6 ELISA Linearity

Scientific Data Images for Mouse IL-6 Quantikine ELISA Kit

Mouse IL-6 ELISA Standard Curve

Mouse IL-6 ELISA Standard Curve

Mouse IL-6 ELISA Kit - Quantikine

Mouse IL-6 ELISA Kit - Quantikine

Increased production of proinflammatory cytokines and VEGF in the plasma and chondrocytes in high fat diet (HFD)-feeding mice.(a~d) levels of plasma IL-6 (a), TNF-alpha (b), IL-1 beta (c) and VEGF (d) in the C57BL/6 male mice fed with a normal diet (ND) or HFD for 8 weeks. (n = 5, **P < 0.01). (e~h) Relative mRNA expression of IL-6 (e), TNF-alpha (f), IL-1 beta (g) and VEGF (h) in the chondrocytes isolated from the 8-week ND or HFD-feeding mice. (n = 5, **P < 0.01). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26271607), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Mouse IL-6 Quantikine ELISA Kit by ELISA

Detection of Mouse Mouse IL-6 Quantikine ELISA Kit by ELISA

PYC suppressed LPS-induced proinflammatory cytokine production in BV2 microglia.Cells were incubated with the indicated concentrations of PYC or vehicle for 1h before LPS treatment (500ng/mL). After 24 h incubation, the culture supernatants were collected, and the amount of TNF-alpha, IL-6, IL-1 beta and IL-10 were measured by ELISA (A-D). Data are represented as mean ± SD from at least 3 independent experiments. *P< 0.05 compared with LPS alone. #P< 0.05. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26367267), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse IL-6

Detection of Mouse IL-6

FFA and lactate enhance production of proinflammatory cytokines and VEGF via HIF1 alpha in mouse chondrocytes.(a) Immunoblotting assay of HIF1 alpha in primary chondrocytes transfected with a scramble siRNA (siNC, 20 nM), a mouse HIF1 alpha specific siRNA1 (siH-1, 20 nM) or siRNA2 (siH-2, 20 nM) for 36 h. (b~e) Relative mRNA levels of IL-6 (b), TNF-alpha (c), IL-1 beta (d) and VEGF (e) in the mouse primary chondrocytes transfected with siNC (20 nM), siH-1 (20 nM) or siH-2 (20 nM)for 12 h plus additional treatment with BSA (5%) or FFA (200 μM) for 24 h. Values not sharing a common superscript letter differ significantly. (n = 3, P < 0.05). (f~i) Relative mRNA levels of IL-6 (f), TNF-alpha (g), IL-1 beta (h) and VEGF (i) in the mouse primary chondrocytes transfected with siNC (20 nM), siH-1 (20 nM) or siH-2 (20 nM) for 12 h plus additional treatment with lactate (25 mM) or vehicle control for 24 h. Values not sharing a common superscript letter differ significantly. (n = 3, P < 0.05). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26271607), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse IL-6

Detection of Mouse IL-6

HFD-induced plasma saturated free fatty acid (FFA) and lactate trigger production of proinflammatory cytokines and VEGF in mouse chondrocytes.(a~d) Relative mRNA levels of IL-6 (a), TNF-alpha (b), IL-1 beta (c) and VEGF (d) in the mouse primary chondrocytes treated with full or fractionated (>3 KD or <3 KD) serum from the 8-week ND or HFD-feeding mice. (n = 3, *P < 0.05, **P < 0.01 and ***P < 0.005). (e~h) Relative mRNA levels of IL-6 (e), TNF-alpha (f), IL-1 beta (g) and VEGF (h) in the mouse primary chondrocytes treated with BSA (5%) or BSA-linked FFA (18:0, 200 μM) for 24 h. (n = 3, *P < 0.05 and **P < 0.01). (i~l) Relative mRNA expression of IL-6 (i), TNF-alpha (j), IL-1 beta (k) and VEGF (l) in the mouse primary chondrocytes treated with lactate (25 mM) or vehicle control for 24 h. (n = 3, *P < 0.05 and **P < 0.01). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26271607), 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-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

Alternate Names

BSF-2, BSF2, IFNB2, IL6, MGI-2A

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)

Gene Symbol

IL6

Additional IL-6 Products

Product Documents for Mouse IL-6 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse IL-6 Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Mouse IL-6 Quantikine ELISA Kit

Customer Reviews for Mouse IL-6 Quantikine ELISA Kit (20)

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  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 09/05/2024
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum and Plasma and Mouse spleenocytes
    Verified Customer | Posted 02/25/2023
  • Mouse IL-6 Quantikine ELISA Kit
    Name: James AHODANTIN
    Sample Tested: EDTA Plasma
    Verified Customer | Posted 11/15/2022
    Mouse IL-6 Quantikine ELISA Kit M6000B
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 10/26/2022
    Mouse serum
    Mouse IL-6 Quantikine ELISA Kit M6000B
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: mouse serum
    Verified Customer | Posted 06/26/2022
    The R2 of the standard curve was more than 0.99. Very stable Kit!
    Mouse IL-6 Quantikine ELISA Kit M6000B
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Plasma
    Verified Customer | Posted 12/05/2021
    Mouse IL-6 Quantikine ELISA Kit M6000B
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Tissue Homogenates
    Verified Customer | Posted 08/31/2021
    Mouse IL-6 Quantikine ELISA Kit M6000B
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Human NIH/3T3 cell line
    Verified Customer | Posted 08/12/2021
    Mouse IL-6 Quantikine ELISA Kit M6000B
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Plasma
    Verified Customer | Posted 05/26/2021
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Tissue Culture Media
    Verified Customer | Posted 04/07/2020
    Mouse IL-6 Quantikine ELISA Kit M6000B
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Mouse skin
    Verified Customer | Posted 02/24/2020
    Mouse IL-6 Quantikine ELISA Kit M6000B
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: EDTA Plasma and Tissue Homogenates
    Verified Customer | Posted 02/20/2020
    Mouse IL-6 Quantikine ELISA Kit M6000B
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 09/14/2019
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Raquel Arroyo
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 04/29/2019
    The information of the kit could included suggested dilutions for serum
    Mouse IL-6 Quantikine ELISA Kit M6000B
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell Culture Supernates
    Verified Customer | Posted 08/06/2018
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: LL/2 mouse Lewis lung carcinoma cell line
    Verified Customer | Posted 04/18/2018
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: mouse pancreatic cancer tissue
    Verified Customer | Posted 04/02/2018
    Mouse IL-6 Quantikine ELISA Kit M6000B
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Brain tissue
    Verified Customer | Posted 09/01/2017
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum and Brain (cortex) tissue
    Verified Customer | Posted 07/19/2017
    Just wanted to let anyone know that the second kit used worked flawless compared to the review I posted above.
  • Mouse IL-6 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum, Liver tissue, Brain (cerebral cortex), choroid plexus and Adipose tissue
    Verified Customer | Posted 07/15/2016
    One plate performed really good but one plate turned out blank. The costumer support were really helpful with diagnosing the error. Attached you find a photo of the plate that didn't work out:
    Mouse IL-6 Quantikine ELISA Kit M6000B

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FAQs for Mouse IL-6 Quantikine ELISA Kit

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  • Q: Assay Diluent RD1-14 contains visible precipitate. Is this acceptable to use?

    A: A small amount of precipitate is expected and acceptable in Assay Diluent RD1-14. Variability has been observed between vials and diluent lots. It may not fully resolubilize but this will not impact assay performance. It is not recommended to filter the precipitate.

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