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Bridge-It® L-Methionine (L-Met) Fluorescence Assay Kit

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Fast and Easy detection of L-methionine levels and enzymes involved in using Methionine as the substrate or as the product!

The Mediomics Bridge-It® L-methionine fluorescence assay method is based on a combination of well-established fluorescence measurement techniques and a new assay platform design that utilizes DNA-binding proteins as biosensors for their respective small molecule coregulators (ligands). The affinity of the DNA sequence-specific MetJ methionine repressor protein for its unique DNA binding site is greatly increased in the presence of its ligand, S-adenosyl methionine (SAM). For this assay, the MetJ consensus sequence was split into two approximately equal DNA “half-sites” with one half fragment labeled with fluorescein and the other half fragment labeled with Oyster® 645 fluorophore3,4. The relative amount of SAM present in a test sample will influence the amount of DNA-MetJ protein complex formation in the assay. When this complex forms, it brings the fluorescence labeled-DNA half-sites into close proximity and causes a measurable change (increase) in fluorescence signal emission that can be readily measured using a microplate reader (wavelength settings: absorption 485 nm; emission 665 nm). The L-methionine is readily converted to SAM by S-adenosylmethionine synthetase in the presence of ATP and Mg2+. L-Methionine concentrations in test samples are determined using an L- methionine standard curve.

The Bridge-It® L-methionine fluorescence assay method exhibits highly desirable performance characteristics including a high signal to background (S/B) ratio, a good linear dynamic range, and, a detection sensitivity of ~5 μM. The total assay time is 45 minutes.

Bridge-It® L-Methionine (L-Met) Fluorescence Assay Kit is:
Mix test sample or standard with the Assay solution and incubate
Read fluorescent signal after 45 minutes of incubation
Assay measures L-Methionine at a lower limit of sensitivity of 5 µM
Assay is adaptable to both low- and high-throughput screening formats

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*384 well format also available

Assay Kit

Product Size:
96 well format