Our lab performs testing for a variety of cytokines and adipokines from commercially available kits. Common examples include TNF-alpha, IL-6, IL-10, IL-1beta, leptin, and adiponectin. Our price list includes the most commonly used testing platforms. In many cases, one can choose between a traditional ELISA or an assay performed with beads on the single molecule counting instrument (SMCxPro). In other cases, the choice may be between an automated platform, traditional ELISA or RIA.
Please note that measuring ranges and volume required can vary a good bit from platform to platform. Basic information about tube type and sample volume is listed on our pricing sheet and in the table below as that may help you decide. However, we are happy to help you find the most appropriate assay for your specific needs. Please reach out to us by phone or email through the button below.
Below are some specifics for cytokine assays we commonly run. These specifications are for human cytokines. Please check with manufacturer for kit availability for other species. ELISA kits are from R&D Systems and SMCxPro are from MilliporeSigma.
Analyte | Method | Sample Type | # Sample Volume (µL) | Manufacturer Stated Reporting Range | * LLOQ actually obtained (pg/mL) |
IL-1 beta HS | SMCxPro | serum or plasma | 100 | 0.20-50 ph/mL | 0.80 |
IL-6 HS | SMCxPro | serum or plasma | 75 | 0.08-50 pg/mL | 0.02 to 0.31 |
IL-6 HS | ELISA | serum or plasma | 100 | 0.2-10 pg/mL | N/A |
IL-6 | ELISA | serum or plasma | 100 | 3.1-300 pg/mL | N/A |
IL-10 HS | SMCxPro | serum or plasma | 100 | 0.10-100 pg/mL | 0.10 to 0.39 |
TNF-a | SMCxPro | serum or plasma | 100 | 0.02-50 pg/mL | 0.1 to 0.4 |
TNF-a | ELISA | cell supernatant (or higher values expected) | 50 | 15.6-1000 pg/mL | N/A |
TNF-a HS | ELISA | serum or plasma | 50 | 0.2-10 pg/mL | N/A |
VEGF-A | SMCxPro | serum or plasma | 25 | 0.20-100 pg/mL | 0.20 to 0.78 |
VEGF-A | ELISA | serum, plasma | 100 | 15.6-1000 pg/mL | N/A |
VEGF-A | ELISA | cell culture supernatant | 200 | 31.3-2000 pg/mL | N/A |
# Sample volume is for one replicate. We ask that you provide enough sample to run two replicates.
Best practices include:
- Use the same sample type throughout your study.
- Sodium heparin is the preferred sample type due to its stable recovery of many different cytokines (1). Other sample types may be acceptable, especially if you are running only one or a few cytokines.
- Process clinical sample as soon as possible (within 1 hour of collection).
- Store at -80°C, not longer than 2 years.
- Avoid multiple freeze-thaws if possible.*
*Note: Cytokines IL-6 and IL-10 seemed to show stability to multiple freeze-thaws in one study (1).
- de Jager W, Bourcier K, Rijkers GT, Prakken BJ, Seyfert-Margolis V. Prerequisites for cytokine measurements in clinical trials with multiplex immunoassays. BMC Immunol. 2009 Sep 28;10:52. doi: 10.1186/1471-2172-10-52. PMID: 19785746; PMCID: PMC2761376.
- Thavasu PW, Longhurst S, Joel SP, Slevin ML, Balkwill FR. Measuring cytokine levels in blood. Importance of anticoagulants, processing, and storage conditions. J Immunol Methods. 1992 Aug 30;153(1-2):115-24. doi: 10.1016/0022-1759(92)90313-i. PMID: 1381403.
- Flower L, Ahuja RH, Humphries SE, Mohamed-Ali V. Effects of sample handling on the stability of interleukin 6, tumour necrosis factor-alpha and leptin. Cytokine. 2000 Nov;12(11):1712-6. doi: 10.1006/cyto.2000.0764. PMID: 11052823.
- Hennø LT, Storjord E, Christiansen D, Bergseth G, Ludviksen JK, Fure H, Barene S, Nielsen EW, Mollnes TE, Brekke OL. Effect of the anticoagulant, storage time and temperature of blood samples on the concentrations of 27 multiplex assayed cytokines – Consequences for defining reference values in healthy humans. Cytokine. 2017 Sep;97:86-95. doi: 10.1016/j.cyto.2017.05.014. Epub 2017 Jun 6. PMID: 28595117.
- Ashworth M, Small B, Oldfield L, Evans A, Greenhalf W, Halloran C, Costello E. The holding temperature of blood during a delay to processing can affect serum and plasma protein measurements. Sci Rep. 2021 Mar 22;11(1):6487. doi: 10.1038/s41598-021-85052-5. PMID: 33753773; PMCID: PMC7985364.
- Graham C, Chooniedass R, Stefura WP, Lotoski L, Lopez P, Befus AD, Becker AB, HayGlass KT. Stability of pro- and anti-inflammatory immune biomarkers for human cohort studies. JTransl Med. 2017 Mar 2;15(1):53. doi: 10.1186/s12967-017-1154-3. PMID: 28253888; PMCID: PMC5335797.
- Gottfried-Blackmore A, Rubin SJS, Bai L, Aluko S, Yang Y, Park W, Habtezion A. Effects of processing conditions on stability of immune analytes in human blood. Sci Rep. 2020 Oct 15;10(1):17328. doi: 10.1038/s41598-020-74274-8. PMID: 33060628; PMCID: PMC7566484.