Molecular Interactions – Assay Kits

Homogeneous Mix-and-Measure Assay Kits for Cancer Immunotherapy

Biochemical High-Throughput Screening of Cancer Immunotherapy Drug Candidates

Activity Screening of Arginase-1 Using Arginase Gold™

Fluorescence Assay Readouts for the Evaluation of Arginase-1 Activity

Catalog No.
Datapoints Contents
NTRC-hARG-1K 1,000 Arginase-1 enzyme, substrate, assay buffer,
detection reagent and reference inhibitor

Key Features

Arginase Gold™ 

Mild reaction conditions
Ambient temperature
Neutral pH
Homogeneous assay format
Few steps before readout, 1.5 hours from start to end
HTS compatible
End-point measurements
Kinetic experiments
Fluorescent readout

Arginase-1 is an enzyme that converts L-arginine into L-ornithine. Arginase-1 is an important drug target for cancer immunotherapy. The Arginase Gold™ assay technology is based on the detection of Arginase-1 activity with the proprietary probe Arginase Gold™. The fluorescence of the probe is quenched upon conversion of L-arginine into L-ornithine, resulting in a decrease of the fluorescent signal. Arginase Gold™ is a homogenous mix-and-measure (‘addition-only’) assay. Arginine is detected by a single incubation step at room temperature. The assay utilizes a fluorescence-based readout and is robust (Z’-factor > 0.6). Progression of the assay can be followed in real time, allowing for kinetic experiments. Arginase Gold™ was especially developed for use in multi-well plates and for high-throughput screening.

The Arginase Gold™ assay technology
Schematic representation of Arginase Gold™ assay. Enzymatic conversion of L-arginine to L-ornithine is allowed to proceed for 60 minutes, after which the progress of the reaction can be measured immediately with fluorescence detection at 510 nm.

Please contact us for custom-based large quantities or to order components separately.

Activity of Tryptophan-Metabolizing Enzymes Using NFK Green™ -GreenScreen™

Fluorescence Assay for the Evaluation of Human and Mouse IDO1 and TDO Activity

Catalog No.
Datapoints Cell-based Biochemical Contents
NTRC-GSCell-1K 1,000   Probe; substrate; IDO1 and TDO reference inhibitors
NTRC-GSCell-10K 10,000   idem
NTRC-hIDO-1K 1,000   Probe; substrate; assay buffer; human IDO1 protein; IDO1 reference inhibitor
NTRC-hIDO-10K 10,000   idem
NTRC-mIDO-1K 1,000   Probe; substrate; assay buffer; mouse IDO1 protein; IDO1 reference inhibitor
NTRC-hTDO-1K 1,000   Probe; substrate; assay buffer; human TDO protein; TDO reference inhibitor
NTRC-hTDO-10K 10,000   idem
NTRC-mTDO-1K 1,000   Probe; substrate; assay buffer; mouse TDO protein; TDO reference inhibitor

Key Features

NFK Green™ 

Mild reaction conditions
Ambient temperature
Neutral pH
Homogeneous assay format
HTS compatible
End-point measurements
Fluorescent readout

Tryptophan metabolism plays an important role in immune modulation and neurodegenerative disease. Tryptophan-metabolizing enzymes are drug targets for cancer immunotherapy and neurodegenerative disease. We have developed NFK Green™ assays for the high-throughput screening of the tryptophan-metabolizing enzymes indoleamine 2,3-dioxygenase (IDO1) and tryptophan dioxygenase (TDO). The NFK Green™ assay works under mild conditions and is suitable for high-throughput screening. The fluorescence assay is based on a highly specific and unique label, developed by NTRC, named NFK Green™. The protocol allows a rapid implementation of the assay for high-throughput screening or regular compound testing.

Schematic representation of NFK Green™ assay
Schematic representation of NFK Green™ assay. NFK Green™ reacts with NFK to form a green fluorescent molecule, with an excitation wavelength of 400 nm and an emission wavelength of 510 nm. At this wavelength, fluorescence interference of library compounds is minimal.

Please contact us for custom-based large quantities or to order components (protein, reagents, and buffers) separately.

References

Grobben et al. (2020) High-throughput fluorescence-based activity assay for Arginase-1. SLAS Discovery, 25 (9):1018-1025.

Seegers et al. (2014) High-throughput fluorescence-based screening assays for tryptophan-catabolizing enzymes. Journal of Biomolecular Screening, 19 (9):1266-1274.