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This standard should be reconstituted in 100 µL Milli-Q® grade water. The reconstituted analyte should be used within 60 minutes or aliquoted into screw-capped polypropylene vials and stored at -20°C for further experiments. Avoid multiple freeze-thaw cycles. One vial contains an amount of analyte sufficient for performing 10 standard curves.
|Assay Target Class||Cytokine|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||0.1 µg|
Pyrogens are substances that cause fever in humans and animals. They can be endogenous or exogenous. One of the characteristics of endogenous pyrogens is their ability to induce cytokine production (IL-1ß, IL-6, IL-8, and TNFa) by the body's immune system. The detection of these pyrogens in pharmaceutical products is an essential regulatory requirement to ensure product quality and patient safety.
In recent years, the Monocyte Activation Test (MAT) has become the most attractive and relevant method for humans. This method enables the detection and quantification of substances with a pyrogenic effect, i.e. those causing a febrile response, and does not require the use of animals.
This application note demonstrates the usefulness of the AlphaLISA® IL-6 kit for the quantification of pyrogens and the ease of use of a homogeneous, wash-free assay with low sample volumes.
Biomarker levels were measured directly in CulturPlates-96 and-384 in a simple, fast, all-in-one-well AlphaLISA assay format. The elimination of transfer and wash steps simplifies cellular assays, reduces variability and significantly reduces hands-on time and costs associated with consumables. Integral plasma membrane (EGFR) and secreted solubleproteins (TNFa, IL1ß, IL6, IL8) were successfully determined, on adherent or suspension cells, using the standard AlphaLISA Immunoassay buffer.
The AlphaLISA® assay is a homogeneous immunoassay alternative to classical ELISA. AlphaLISA assays were originally utilized to detect analytesin cell cultures upernatants or serum/plasma samples.