1. HTRF® cytokine assays

2. Cytokine Assays
Applications:
Cytokine quantification
Readout of kinase activity or receptor activation
Infectious disease
Allergy studies
Autoimmune disorders
Streamlined protocols for cytokine measurement directly on cells or cell supernatant
Sandwich immunoassay involving two monoclonal antibodies for batch to batch reproducibility
Highly sensitive (< 15 pg/mL)
Small sample size needed (down to 5µl)
Adaptable to HTS
No washing step and much lower sample volumes than traditional ELISA, making it both cost and time efficient
HTRF biomarkers

3. A comprehensive line of ready-to-use kits for quantifying cytokines in cell supernatant or whole cell
Comprehensive reagent supply: calibrator, conjugates, assay buffers and complete protocol
Small and bulk size tests
All batches systematically controlled
Assays available for:
(m&h) TNFa, (h) IL1b, (h) IL2, (h) IL4, (h) IL5, (m&h) IL6, (h) IL 8, (h)IL10, (m&h) IL17, (h) IL12, (m&h) IFNg, (h) GROa
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HTRF biomarkers

4. Cytokine assays can be run using cell supernatants or whole cells
Sandwich assays using a mix-and-measure procedure
HTRF biomarkers

5. Cell-based and supernatant formats give similar results
Not sure if we want the individual pages cytokines, PGE2 and Histamine to be more academic with bullets describing theory, etc. or if to leave more assay focused??? Slide text…. Cisbio offers kits for cytokine detection to address the screening of major inflammation targets
HTRF biomarkers

6. Product specifications (1/3)
(h) TNF alpha 
(m) TNF
alpha
(h) IL1b 
(h) IL2 * 
(h)IL4
(h)IL5
Assay type:
Sandwich 
Sandwich
Detection limit 
10 pg/mL 
5 pg/mL
15 pg/mL 
39 pg/mL 
16 pg/mL
39 pg/mL
Working range
Up to 6000 pg/mL
Up to pg/mL
Up to 5000 pg/mL
Up to 10,000 pg/mL
Incubation
3-24h at RT
3h at RT
2h at RT
Diluent
Phosphate buffer, culture medium
* Assay on demand (frozen, available in 1,000 and 20,000 test volumes)
HTRF biomarkers 6

7. Product specifications (2/3)
(h) IL6 
(m) IL6 
(h) IL8 * 
(h) IL10 
(h) IL12 
Assay type:
Sandwich 
Detection limit 
15 pg/mL 
40 pg/mL 
15 pg/mL 
10 pg/mL 
30 pg/mL 
Working range
Up to pg/mL
Up to 5000 pg/mL ND
Incubation
3h at RT
2h at RT
ON at 4C
4h at RT
Diluent
Phosphate buffer, culture medium
* Assay on demand (frozen, available in 1,000 and 20,000 test volumes)

8. Product specifications (3/3)
(h) IL17 
(m) IL17*
(h) IFNg
(m) IFNg
(h) GROa
Assay type:
Sandwich 
Detection limit 
56 pg/mL 
10 pg/mL 
30 pg/mL 
19 pg/mL 
Working range
Up to 5000 pg/mL
Up to 2000 pg/mL NA
Incubation
24h at RT
2h at RT
ON at RT
2h at 4°C
Diluent
Phosphate buffer, culture medium
* Assay on demand (frozen, available in 1,000 and 20,000 test volumes)

9. Learn how they used the kits
TNF-alpha Krinstin P et al., «  Two High Throughput Screen Assays for the Measurement of TNF-alpha in THP-1 Cells », Current chemical Genomics, 2011, 5,
IFN-gamma K Enamoto et al., «  High throughput screening for human interferon-gamma production inhibitor using homogenous time-resolved fluorescence. », J Biomol Screen Aug;5(4):263-8
Cytokines S Achardet al., «  Homogeneous assays allow direct "in well" cytokine level quantification. », Assay Drug Dev Technol Feb;1(1 Pt 2):181-5
HTRF biomarkers