Product Overview

The PeliKine Human IL-6 ELISA Rapid Kit is the optimal tool for rapid, sensitive and specific detection of the human pro-inflammatory cytokine Interleukin 6 (IL-6) in the PyroCell^® Monocyte Activation Test (MAT). IL-6 cytokines are released by human monocytes during exposure to product samples containing endotoxins, or pyrogens. IL-6 is a mediator of inflammatory responses in humans and is involved in the induction of acute phase proteins [1-4] and the development of fever [5]. 

The PeliKine Human IL-6 ELISA Rapid Kit contains all necessary reagents to conduct a "sandwich-type" of enzyme-linked Immunosorbent Assay (ELISA). Secreted human IL-6 cytokines contained in the cell culture supernatants during Monocyte Activation are simultaneously bound to anti-IL-6 capture antibodies coated to the polystyrene microtiter wells of a 96-well plate, and biotinylated anti-IL-6 antibodies added in the same incubation step. Unbound material is removed by washing. Subsequently, a horseradish peroxidase (HRP) conjugated streptavidin is added that binds onto the biotinylated side of the cytokine sandwich. After removal of unbound material by washing, a substrate solution is added.  

A colored product is formed in proportion to the amount of IL-6 from MAT cell culture supernatants, or IL-6 reference standard dilutions. After the reaction is terminated by adding the stop solution, the absorbance (optical density, OD) is measured in a microtiter plate reader at 450nm and a reference wavelength (540 to 590nm).  

The PeliKine Human IL-6 ELISA Rapid Kit has been validated for use with PyroCell^® MAT Kit and the PyroCell^® MAT Human Serum Kit containing a cell preparation with naïve monocytes and a serum source for conducting the monocyte activation test. Together, they represent the PyroCell^® MAT Assay System and the PyroCell^® MAT HS System, respectively each supports the testing of up to nine product samples according to the compendial requirements of the European Pharmacopeia (EP 2.6.30). The sensitivity of the assay system is <1pg/mL. 

The Monocyte Activation Test is performed in two steps: during an overnight cell culture step, a monocytic cell preparation, e.g. peripheral blood mononuclear cells (PBMC) is stimulated for release of inflammatory cytokines with a product preparation containing a pyrogenic contaminant. On the next day, the released cytokines are detected with an ELISA assay. The color reaction is measured  with an absorbance readers, e.g. the Nebula^® Absorbance Reader or the Nebula Multimode Reader, at a wavelength of 450 nm and a reference wavelength of 540-590 nm. The pyrogenic content of the sample is then analyzed against the reference standard endotoxin (RSE) and is expressed in Endotoxin Equivalent Units (EEU/mL).

The PeliKine Human IL-6 ELISA Rapid Kit is the new generation of the traditional PeliKine compact human IL-6 ELISA.  And will replace the traditional ELISA (Part # 253325) and will fully replace it by early 2024. For more information please contact our Scientific Support Team. 

References

1. Gauldie,J. et al (1987) Proc.Natl.Acad.Sci.(USA) 84: 7251
2. Le,J. and Vilcek,J. (1989) Lab.Invest. 61: 588
3. Heinrich,P.C. et al (1990) Biochem.J. 265: 621
4. Kishimoto,T. (1989) Blood 74: 1
5. Helle,M. et al (1988) Eur.J.Immunol. 18: 957