Disturbances in cytokine regulation are known to lead to inappropriate or ineffective immune responses. Cytokines are of particular interest in the fields of oncology, infectious diseases, allergies and autoimmune diseases. Cisbio Bioassays has developed a line of reagents for quantifying a variety of cytokines, based on HTRF® technology. These reagents produce quality results, are highly sensitive and can be used with cell supernatants or whole cells.
All cytokine assays can be run with cell supernatants or whole cells using the protocols shown here. Cytokines are measured using sandwich immunoassays involving two monoclonal antibodies: anti-IL1β (MAb1) labeled with Eu-Cryptate and anti-IL1β (MAb2) labeled with XL665. These antibodies may be pre-mixed and added in a single dispensing step, to further streamline the protocol. In addition, we have adapted our assay protocols to incubate the labeled antibody pair with cells during the stimulation step, and thus eliminate the need to transfer cell supernatants prior to detection. The addition of potassium fluoride stops cell stimulation and allows the HTRF signal to be read immediately.
| Incubation | 2 h at RT |
|---|---|
| Calibration range: | 0-2,000 pg/mL |
| Detection limit | < 15 pg/mL NIBSC 89/680 |
| Extended range | Up to 10,000 pg/mL |
Cisbio Bioassays's assays are ideal for highly sensitive cytokine screening. They have rapid, easily miniaturized protocols that can be run in a single plate, from cell stimulation through to assay measurement. Figure 1 shows levels of TNFα (panel A), IFNγ (panel B), IL1β (panel C) and IL6 (panel D) detected using the cell-based and supernatant assay formats. These data show that both protocols give similar results and consistent cytokine concentrations with respect to the number of cells per well, indicating that the presence of cells does not alter assay performance. Levels of IL6 (panel D) were not quantified above 6,250 cells per well, as all concentrations were off the scale.
| Description | Cat. no | Product insert | MSDS |
|---|---|---|---|
| Human IL1 beta kit - 1,000 tests | 62IL1PEB | ||
| Human IL1 beta kit - 20,000 tests | 62IL1PEC |
| Description | Cat. no | Product insert | MSDS |
|---|---|---|---|
| Diluent 1 - 20 ml | 62DL1DDD | - | |
| Human IL1 beta calibrator | 62IL1CDA | - | |
| Human IL1 beta control | 62IL1TDA | - | |
| Reconstitution buffer 2 - 13 ml | 62RB2RDD | - | - |
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