LumiMAT™ Pyrogen Detection Kit

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

298-36991

LumiMAT™ Pyrogen Detection Kit - Cells

96 tests (-80°C)

297-96801

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

*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.

LumiMAT Fig 1

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.

LumiMAT Fig 2