LumiMAT™ Pyrogen Detection Kit

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This kit is for Monocyte Activation Test (MAT) to detect pyrogens. Pyrogen is a generic term for substances that cause body temperature rise in animals. Parenteral drugs and medical devices that contact to blood must be free from pyrogens. The first developed test to detect a wide range of pyrogens was Rabbit Pyrogen Test (RPT). In RPT, samples to be tested are intravenously administered to rabbits and monitor the body temperature. Since RPT has problems with reproducibility, accuracy, and cost, Limulus Amebocyte Lysate (LAL) reagent has beem widely used as the replacement. However, LAL reagent cannot detect non-endotoxin pyrogens, and RPT is still used in case LAL reagent is not appropriate.

MAT is an in vitro pyrogen test developed as an alternative of RPT and it can detect not only endotoxin but also non-endotoxin pyrogens.

KEY FEATURES

• Assay with reporter NOMO-1 cells
• Rapid testing (5 hours)
• Convenient reporter assay
• Only add luminescent substrate to the plate after incubation
• Stable reporter cell line
• Cell lines stably expressing reporter genes
• High reproducibility with less lot-to-lot variation
• Stable supply from master cell bank

KIT COMPONENTS

*BothLumiMAT™ Pyrogen Detection Kit - Cells and Reagent Set are necessary for an assay.

ASSAY PRINCIPLE

MAT is used to detect pyrogens that activate human monocytic cells to release inflammatory cytokines.

Conventional MAT

Culture PBMCs (peripheral blood mononuclear cells) and samples to be tested in microwell plate. Cytokines released from monocytic cells by exposure to pyrogens are measured by ELISA to detect pyrogens in the samples.

LumiMAT™

LumiMAT™ uses a monocytic cell line NOMO-1 in which a luciferase reporter gene is introduced to express luciferase protein in response to NF-kB signals activated by exposure to pyrogens. Because the expressed luciferase generates luminescence by reacting with the substrate, luminescence can be detected by a luminescence microplate reader to detect pyrogens in the samples. 

ASSAY FLOW

Add samples to be tested or standard endotoxin into microplate and add reporter cells and incubate for 3 hours.After the incubation add luminescent substrate tomeasure luminescence with a plate reader. Compared to ELISA, this is a simple one-plate assay that suppresses inter-test variations. In addition, results can be obtained in about 5 hours, which is significantly shorter than the 1.5 days required by the conventional method.