Cytokine Receptor Assays

Functional Assays for >85% of Human Interleukins and Their Receptors

Cytokine receptors are critical for cellular and extracellular communication and mediating immunological responses. These receptors include type 1 and 2 cytokine-like interleukins, GM-CSF, interferons as well as chemokines, TNFα, TGF-β, and Ig superfamily receptor types. All cytokine receptors are associated with one or more members of JAKs, which couple ligand binding to tyrosine phosphorylation of various signaling proteins (STATs) recruited to the receptor complex.

Eurofins DiscoverX offers a comprehensive portfolio of cell-based assays to enable the study and targeting of human cytokine proteins and their receptors. These assays are designed to be highly specific and reproducible, include a simple protocol, and have a large signal-to-noise ratio and matrix tolerance. These assay qualities have enabled their broad usage in functional screening, functional characterization, QC lot release assays, and neutralizing antibody studies.

New! Visit discoverx.com/covid-19 or click on the Video link below to learn about therapeutics targeting proinflammatory cytokines associated with COVID-19 disease progression.
 
View Video  

Advantages of Cytokine Assays

  • Biologically-Relevant — MOA-reflective, functional assays for monitoring cytokine receptor activation and dimerization 
  • Qualified Bioassays – Accelerate your drug release program into QC lot release with bioassays qualified using approved therapeutics like Actemra® (tocilizumab), Kineret® (anakinra), Leukine® (sargramostim), and Humira® (adalimumab)
  • Easy-to-Run & Scalable – Simple, rapid, and homogeneous protocol amenable to high-throughput formats for increased efficiency​

Key Resources

  • Knowledge-Based Video: Accelerate Your COVID-19 Drug Discovery with Qualified Cell-Based Assays for Pro-inflammatory Cytokines 
  • Related Product Pages
    • Bioassays – Ready-to-use cell-based assays for potency measurement from characterization to  QC lot release testing of biologics
    • Receptor Kinase Assays – Receptor tyrosine kinase and cytokine receptor assays associated with SH2 recruitment and JAK1, JAK2 and JAK3 kinases
    • Signaling Pathway Reporter Assays – Cell lines to quantify the activation or inhibition of signaling pathways with a downstream read-out 
    • COVID-19 – Functional cell-based assays to support your COVID-19 drug discovery through QC lot release programs
    • Targeted Protein Degradation – Assays for screening of small molecule therapeutics and quantification of changes in endogenous protein levels in disease-relevant cell models
    • GPCR Assays – S1P (sphingosine-1-phosphate) and the LPA (lysophosphatidic acid) subfamilies, C5AR1, and other GPCR assays for analyzing cAMP accumulation, calcium flux, β-arrestin recruitment, and receptor internalization

 

Eurofins DiscoverX cytokine receptor assays are available as complete ready-to-use bioassay and eXpress kits and stable cell lines formats for receptor activation and dimerization assessment. Please contact custom assay development for additional assays. Learn more about the updated Cell Culture and Handling Procedure in this Technical Bulletin.

Assays for Cytokines Receptors

Ligand Assay Mechanism Cell Line Bioassay Kit
(2- & 10-plate)
eXpress Kit
(2- & 10-Plate)
Ligand Synonyms
CLCF1 - Contact Us     CLCF1; BSF-3; BSF3; CISS2; CLC; NNT-1; NNT1; NR6
CNTF - Contact Us     CNTF ; HCNTF
CTF1 - Contact Us     CTF1
G-CSF SH2 Recruitment 93-0809C3   93-0809E3CP16M
93-0809E3CP16L
CSF3; G-CSF; GCSF
GM-CSF CSFR2A / CSFR2B 93-1078C3     CSF2; GMCSF
IL-1α IkB degradation 93-0538C15     IL1A; IL-1A; IL1; IL1-ALPHA; IL1F1
IL-1α IL-1R1 / IL-1RAP 93-1032C3 93-1032Y3-00105
93-1032Y3-00106
93-1032E3CP5M
93-01032E3CP5L
IL1A; IL-1A; IL1; IL1-ALPHA; IL1F1
IL-1β IkB degradation 93-0538C15     IL1B; IL-1; IL1-BETA; IL1F2
IL-1F10 - Contact Us     IL1F10; FIL1-theta; IL-1HY2; IL-38;
IL1-theta; IL1HY2
IL-1RN IkB degradation 93-0538C15     IL1RN; DIRA; ICIL-1RA; IL-1RN;
IL-1ra; IL-1ra3; IL1F3; IL1RA; IRAP; MVCD4
IL-1RN IL-1R1 / IL-1RAP 93-1032C3 93-1032Y3-00105
93-1032Y3-00106
93-1032E3CP5M
93-1032E3CP5L
IL1RN; DIRA; ICIL-1RA; IL-1RN; IL-1ra; IL-1ra3; IL1F3; IL1RA; IRAP; MVCD4
IL-2 IL-2RB / IL-2RG 93-0998C3   93-0998E3CP5M
93-0998E3CP5L
IL2; IL-2; TCGF; lymphokine
IL-2 IL-2RB / IL-2RG / IL-2RA 93-1003C3 93-1003Y3-00091
93-1003Y3-00092
93-1003E3CP0M
93-1003E3CP0L
IL2; IL-2; TCGF; lymphokine
IL-3 IL-3R / CD131 (CSF2RB) 93-0969C1     IL3; IL-3; CD131; CD-131
IL-4 IL-4R / IL-2RG 93-0988C3   93-0988E3CP5M
93-0988E3CP5L
IL4; BCGF-1; BCGF1; BSF-1; BSF1; IL-4
IL-4 IL-4R / IL-13R 93-1000C3
93-1001C1
  93-1000E3CP5M
93-1000E3CP5L

93-1001E1CP0M
93-1001E1CP0L
IL4; BCGF-1; BCGF1; BSF-1; BSF1; IL-4
IL-5 IL-5R / CSF2RB 93-0972C3   93-0972E3CP0M
93-0972E3CP0L
IL5; EDF; IL-5; TRF
IL-6  IL-6RA / IL-6ST 93-1045C3 93-1045Y3-00043
93-1045Y3-00044
93-1045E3CP5M
93-1045E3CP5L
IL6; BSF2; HGF; HSF; IFNB2; IL-6
IL-7 IL-7R / IL-2RG 93-0997C13 93-0997Y13-00081
93-0997Y13-00082
93-0997E13CP0M
93-0997E13CP0L
IL7; IL-7
IL-8 CXCR2  93-0202C2  

93-0202E2CP2M

93-0202E2CP2L

CXCL8; GCP-1; GCP1; IL8; LECT; LUCT; LYNAP; MDNCF; MONAP; NAF; NAP-1; NAP1
IL-8 CXCR1 93-0226C3   93-0226E3CP0M
93-0226E3CP0L
CXCL8; LGCP-1; GCP1; IL8; LECT; LUCT; LYNAP; MDNCF; MONAP; NAF; NAP-1; NAP1
IL-9 IL-9R / IL-2RG 93-1036C1     IL9; HP40; IL-9; P40
IL-10 IL-10RA / IL-10RB 93-0985C3 93-0985Y3-00135
93-0985Y3-00136
93-0985E3CP0M
93-0985E3CP0L
IL10; CSIF; GVHDS; IL-10; IL10A; TGIF
IL-11 IL-11R / IL-6ST 93-1028C1     IL11; AGIF; IL-11
IL-12 IL-12RB1 / IL-12RB2 93-1041C3   93-1041E3CP0M
93-1041E3CP0L
IL12, p70
IL-13 IL-4R / IL-13R 93-1000C3   93-1000E3CP5M
93-1000E3CP5L
93-1001E1CP0M
93-1001E1CP0L
IL13; IL-13; P600
IL-13 IL-4R / IL-13R 93-1001C1   93-1001E1CP0M IL13; IL-13; P600
IL-14 - Contact Us     TXLNA; IL14; TXLN
IL-15 IL-2RB / IL-2RG 93-0998C3   93-0998E3CP5M
93-0998E3CP5L
IL15; IL-15
IL-16 - Contact Us     IL16; LCF; NIL16; PRIL16; prIL-16
IL-17A IL-17RA / IL-17RC 93-0999C3
93-0931C1
93-0999Y3-00053
93-0999Y3-00054
93-0999E3CP0M
93-0999E3CP0L
93-0931E1CP7M
93-0931E1CP7L
IL17A; CTLA8; IL-17; IL-17A; IL17
IL-17B - Contact Us      
IL-17C - Contact Us      
TNFα IL-17RD / TNFR2 93-1068C3   93-1068E3CP5M
93-1068E3CP5L
TNF-alpha
IL-17E IL-17RA / IL-17RB 93-1105C3     IL25; IL17E
IL-17F IL-17RA / IL-17RC 93-0999C3   93-0999E3CP0M
93-0999E3CP0L
IL-17F
IL-18 IL-18R / IL-18RAcP 93-1131C3     IL18; IGIF; IL-18; IL-1g; IL1F4
IL-20 IL-20R1 / IL-20R2 93-1027C3   93-1027E3CP0M
93-1027E3CP0L
IL20; IL-20; IL10D; ZCYTO10
IL-20 IL-22R / IL-20R2 Contact Us     IL20; IL-20; IL10D; ZCYTO10
IL-21 IL-21R / IL-2RG 93-1035C3   93-1027E3CP0M
93-1035E3CP0L
IL21; CVID11; IL-21; Za11
IL-22 IL-22R / IL-10RB  93-1026C1   93-1026E1CP7M
93-1026E1CP7L
IL22; IL-21; IL-22; IL-D110; IL-TIF;
ILTIF; TIFIL-23; TIFa; zcyto18
IL-23 IL-23R / IL-12RB1 93-1007C3   93-1007E3CP0M
93-1007E3CP0L
IL23A; IL-23; IL-23A; IL23P19; P19;
SGRF
IL-24 IL-20R1 / IL-20R2 93-1027C3     IL24; C49A; FISP; IL10B; MDA7; MOB5; ST16
IL-24 IL-22R / IL-20R2 Contact Us     IL24; C49A; FISP; IL10B; MDA7; MOB5; ST16
IL-26 IL-20R1 / IL-10RB 93-1020C3   93-1020E3CP0M
93-1020E3CP0L
IL26; AK155; IL-26
IL-27 IL-27RA / IL6ST 93-1119C3     IL30, p28
IL-28A IFNLR1 / IL-10RB 93-0993C3    93-0993E3CP5M
93-0993E3CP5L
IL28A, IFNL2
IL-28B IFNLR1 / IL-10RB 93-0993C3    93-0993E3CP5M
93-0993E3CP5L
IL28B, IFNL3
IL-29 IFNLR1 / IL-10RB 93-0993C3    93-0993E3CP5M
93-0993E3CP5L
IFNL1; IL-29; IL29
IL-31 OSMRb / IL-31RA 93-1002C3 93-1002Y3-00083
93-1002Y3-00084
93-1002E3CP5M
93-1002E3CP5L
IL31; IL-31
IL-32 - Contact Us     IL32; IL-32alpha; IL-32beta;
IL-32delta; IL-32gamma; NK4; TAIF; TAIFa; TAIFb; TAIFc; TAIFd
IL-33 IL-1RL1 (ST2) / IL-1RAP 93-1067C3 93-1067Y3-00079
93-1067Y3-00080
93-1067E3CP5M
93-1067E3CP5L
IL33; C9orf26; DVS27; IL1F11;
NF-HEV; NFEHEV; RP11-575C20.2
IL-34 CSF1R / CSF1R 93-1061C3     IL34; C16orf77; IL-34
IL-35 IL-12RB2 / IL-6ST Contact Us     IL-35
IL-36A IL-1RL2 / IL-1RAP Contact Us     IL36A; FIL1; FIL1(EPSILON); FIL1E; IL-1F6; IL1(EPSILON); IL1F6
IL-36B IL-1RL2 / IL-1RAP Contact Us     IL36B; FIL1; FIL1-(ETA); FIL1H; FILI-(ETA); IL-1F8; IL-1H2; IL1-ETA; IL1F8; IL1H2
IL-36C IL-1RL2 / IL-1RAP Contact Us     IL36G; IL-1F9; IL-1H1; IL-1RP2; IL1E;
IL1F9; IL1H1; IL1RP2
IL-36RN IL-1RL2 / IL-1RAP Contact Us     IL-36RA; IL36RA
IL-37 IL-18R / SIGIRR Contact Us     IL37; FIL1; FIL1(ZETA); FIL1Z; IL-1F7; IL-1H; IL-1H4; IL-1RP1; IL-37; IL1F7; IL1H4; IL1RP1
LIF IL-6ST / LIFR Contact Us     LIF; CDF; DIA; HILDA; MLPLI
NP - Contact Us     Neuropoietin
M-CSF CSF1R / CSF1R 93-1061C3     CSF1; CSF-1; MCSF
OSM OSMRb / IL-6ST 93-1056C3     OSM; MGC20461
TSLP IL-7R / TSLP-R 93-1019C13   93-1019E13CP25M
93-1019E13CP25L
TSLP

Eurofins DiscoverX cytokine receptors are based on the Enzyme Fragment Complementation (EFC) technology. These assays allow for the analysis of receptor activation and dimerization assessment through various assay modalities to ultimate allow for screening, potency and stability testing of therapeutic candidates.  Select assay principles for receptor dimerization and IB protein degradation assays are shown below. For additional assay principles, please refer to receptor kinase, signaling pathway reporter, targeted protein degradation, and GPCR landing pages.
 


PathHunter® Tocilizumab Bioassay Assay Principle

12022-(1).png
Measure IL-6 induced heterodimerization of IL-6 receptor (IL-6R) with the IL-6 Signal Transducer protein (IL-6ST; gp130) using the PathHunter Tocilizumab Bioassay (Cat. No. 93-1045B3-00110) qualified with the FDA approved therapeutic Actemra (tocilizumab, an anti-IL-6R antibody being evaluated as potential therapeutic for COVID-19). The assay principle involves the complementing of enzyme donor ProLink™ (PK) and enzyme acceptor (EA) EFC fragments tagged to IL-6ST and IL-6R, respectively. In presence of IL-6, the receptor units dimerize and there is an increase in signal detected upon substrate addition. However, in the presence of tocilizumab, the binding of IL-6 is inhibited and the signal decreases. Actemra® a registered trademark of Chugai Seiyaku Kabushiki Kaisha Corp., a member of the Roche Group.
 


PathHunter Adalimumab Bioassay Assay Principle

11807-(1).png
Detect IkB protein degradation as a result of TNFα-mediated activation of the NF-kB signaling pathway using PathHunter Adalimumab Bioassay Kit (Cat. No. 93-0538B15-00132) qualified with the FDA approved therapeutic Humira (adalimumab), which is being evaluated as potential therapeutic for COVID-19. The assay principle involves the enzyme donor ProLabel® (PL) tagged to phosphorylated IkB, and the IκB levels can then be measured by the addition of EA, which forces complementation of the two EFC fragments. The resulting active enzyme hydrolyzes the substrate to generate the signal that is proportional to the degree of IκB stabilization. In presence of TNFα, which activates the NF-kB signaling pathway,  proteasome-mediated IκB degradation occurs and there is a loss of signal. In the presence of both TNFα and adalimumab, which blocks the binding of TNFα, degradation of IκB is avoided, and the EFC complementation occurs leading to an increase in signal upon substrate addition.  Humira® is a registered trademark AbbVie, Inc.
 

Analyze Multiple Interleukin Receptor Famillies with Highly Specific Assays

12027-(1).png

Representative examples of assays for interleukin receptors from 6 different interleukin receptor families. Each plot shows a dose response for the relevant ligand(s) in a given assay from the indicated family. Data plotted are mean RLU and standard deviation from at least triplicate wells for each dose. These assays are characterized by robust assay windows and low CVs.

 

Qualify Therapeutics with MOA-Reflective, Receptor Dimerization Bioassays

12025-(3).png
 

The PathHunter Anakinra Bioassay (Cat. No. 93-1032Y3-00106) was used to quantify an inhibitory response from Kineret (anakinra, an IL-1R antagonist being evaluated as potential therapeutic for COVID-19). [A.] In the presence of anakinra, there is a dose dependent inhibition (signal-to-background (S/B) 4.2; IC50 302 ng/mL) of IL-R1 and IL-1RAP dimerization. In comparison with agonist IL-1β, the receptors dimerize and there is an increase in signal detected (S/B 5.3; EC50 4.2 ng/mL). [B.] Dilutional linearity derived from the relative potency experiments performed over a range of 50-150%. Overall, the bioassay measures a robust response to both anakinra and IL-1β with a high accurate potency of over 99.4%. Kineret® is a registered trademark, licensed by Swedish Orphan Biovitrum AB and marketed by Sobi, Inc.

 

Accurately Measure Receptor Activation with Highly Reproducable Bioassays for QC Lot Release

12028-(1).png

Tocilizumab Bioassay (Cat. No. 93-1045B3-00110) assessment of intra-day plate-to-plate and inter-day variability. Three experimental replicates of the assay were prepared as 11-point dose-response curves and run on the same plate with duplicate wells per dose on the same day by one operator. Results indicate 978, 960 and 997 ng/mL mean IC50’s for plates 1-3, respectively, and an overall 9.7% intermediate precision IC50. Similar experiments were repeated again to obtain data from three individual days (data not shown) for inter-day variability assessment with results showing an overall 1330 ng/mL mean IC50 and 24.6% intermediate precision IC50 for the 3 days. Results from both experiments for inter-plate and inter-day variability analysis show excellent uniformity demonstrating high reproducibility required for bioassay implementation in a QC environment for drug release.