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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.
RPT
LAL
MAT
Pyrogen
Endotoxin (derived from Gram-negative bacterial cell walls)
✔
✔
✔
Non-endotoxin Pyrogens (derived from Gram-positive bacteria, viruses, fungi etc.)
✔
Not-detectable
✔
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
Catalog Number
Product Name
Contents
283-42599
LumiMAT Pyrogen Detection Kit - Cells
96 tests (-80°C)
286-42609
LumiMAT Pyrogen Detection Kit - Reagent Set
96 tests (-20°C)
Kit Components
1
LumiMAT Pyrogen Detection Kit - Reagent Set
Assay Medium
20 mL
Dilution Medium
100 mL
Luciferase Assay Buffer
12 mL
Luciferase substrate
240 μL
2
LumiMAT Pyrogen Detection Kit - Cells
LumiMAT Cells
350 μL
*Both LumiMAT 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.
