Indoleamine 2,3-Dioxygenase 1 (IDO1) Inhibitor Assay Kit (ab273304) enables rapid screening of test compounds for modulation of IDO1 activity.
Fluorescent
Quantitative
Application | Reactivity | Dilution info | Notes |
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Application FuncS | Reactivity Reacts | Dilution info - | Notes - |
Catalyzes the first and rate limiting step of the catabolism of the essential amino acid tryptophan along the kynurenine pathway (PubMed:17671174). Involved in the peripheral immune tolerance, contributing to maintain homeostasis by preventing autoimmunity or immunopathology that would result from uncontrolled and overreacting immune responses (PubMed:25691885). Tryptophan shortage inhibits T lymphocytes division and accumulation of tryptophan catabolites induces T-cell apoptosis and differentiation of regulatory T-cells (PubMed:25691885). Acts as a suppressor of anti-tumor immunity (PubMed:23103127, PubMed:25157255, PubMed:14502282, PubMed:25691885). Limits the growth of intracellular pathogens by depriving tryptophan (PubMed:25691885). Protects the fetus from maternal immune rejection (PubMed:25691885).
IDO-1, INDO, IDO, IDO1
Indoleamine 2,3-Dioxygenase 1 (IDO1) Inhibitor Assay Kit (ab273304) enables rapid screening of test compounds for modulation of IDO1 activity.
IDO-1, INDO, IDO, IDO1
Fluorescent
Quantitative
Microplate
Blue Ice
-20°C
-20°C
-20°C
Indoleamine 2,3-Dioxygenase 1 (IDO1) Inhibitor Assay Kit (ab273304) enables rapid screening of test compounds for modulation of IDO1 activity.
Unlike conventional colorimetric IDO1 assays based upon UV absorbance, the assay uses a fluorogenic developer that selectively reacts with NFK to produce a highly fluorescent product detectable in the visible range (Ex/Em = 402/488 nm).
This ensures a high signal-to-background ratio and little interference due to short wavelength absorbance by test compounds. The assay is highly sensitive, has a simple no-wash protocol and is high-throughput adaptable. The kit contains a complete set of reagents sufficient for performing 100 reactions in a 96-well plate format.
This product is manufactured by BioVision, an Abcam company and was previously called K976 Indoleamine 2,3-Dioxygenase 1 (IDO1) Inhibitor Screening Kit. K976-100 is the same size as the 100 test size of ab273304.
This supplementary information is collated from multiple sources and compiled automatically.
Indoleamine 2 3-dioxygenase (IDO) also known as IDO1 is an enzyme involved in the catabolism of tryptophan to kynurenine. It weighs approximately 45 kDa and it is an important target in immunoregulatory processes. IDO is highly expressed in antigen-presenting cells such as dendritic cells and macrophages as well as in various tumor cells where it helps modulate immune responses. This enzyme can also be measured by using assays like IDO ELISA.
IDO plays a role in immune response regulation by degrading tryptophan an essential amino acid needed for T-cell proliferation. The depletion of tryptophan and the accumulation of its metabolites such as kynurenine cause immunosuppressive effects within the tissue microenvironment. While IDO functions mostly as a standalone enzyme its activity influences the cellular surroundings by altering the balance of local immune reactions.
IDO integrates into the tryptophan metabolism pathway and is important in the kynurenine pathway. Through this pathway it maintains immune homeostasis and cell defense mechanisms. Proteins like kynureninase and kynurenine 3-monooxygenase also participate in the same metabolic processes which further extend the effects of tryptophan metabolism on immune cell behavior.
IDO is linked with cancer and chronic inflammatory conditions. In cancer elevated IDO expression suppresses anti-tumor immunity aiding tumor cells to evade immune surveillance. It also plays a role in autoimmune disorders by regulating excessive immune activity. During cancer progression IDO often works in conjunction with immune checkpoint proteins like PD-L1 which further enhances its immunosuppressive capabilities within the tumor microenvironment.
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Example dose response curve.
Example Data
Dose-response curve for IDO1 inhibition by the included selective IDO1 Inhibitor IDO5L. The IC50 value (24.2 ± 1.3 nM) was derived by 4-parameter logistic curve fitting with each point representing the mean ± EM of at least 3 replicates. All assays were performed according to the kit protocol.
Measurement of IDO1 inhibition in presence and absence of 1 μM of IDO5L.
Example Data
Measurement of IDO1 inhibition in presence and absence of 1 μM of IDO5L (a potent, highly selective competitive IDO1 inhibitor) and 100 μM of norharmane (a natural product that acts as a weak IDO1 inhibitor). The no inhibitor reaction contained assay buffer with 0.1% DMSO (v/v) as a solvent control.
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