Indoleamine 2,3-Dioxygenase 1 (IDO1) Activity Assay Kit (ab235936)
Key features and details
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Cell culture extracts, Tissue Extracts
Overview
-
Product name
Indoleamine 2,3-Dioxygenase 1 (IDO1) Activity Assay Kit
See all Indoleamine 2, 3-dioxygenase kits -
Detection method
Fluorescent -
Sample type
Cell culture extracts, Tissue Extracts -
Product overview
Indoleamine 2,3-Dioxygenase 1 (IDO1) Activity Assay Kit (ab235936) enables IDO1 activity to be easily determined in mammalian tissues and cell lines. The assay uses a fluorogenic developer that selectively reacts with N-formylkynurenine (NFK) to produce a highly fluorescent product (Ex/Em = 402/88 nm), ensuring a high signal-to-background ratio. The kit also includes a highly selective IDO1 inhibitor for verification of enzyme activity in biological matrices. The assay has a simple no-wash protocol, is high-throughput adaptable and can detect down to 0.2 mU of IDO1 activity or 200 pmole NFK.
-
Notes
Indoleamine 2,3-Dioxygenase 1 (IDO1, EC 1.13.11.52) is a cytoplasmatic hemoprotein that oxidizes tryptophan yielding N-formylkynurenine (NFK). In mammals, this reaction is the first and rate limiting step in the kynurenine catabolic pathway. IDO1 activity is low under normal physiological conditions, but is dramatically upregulated by proinflammatory cytokines such as interferon-γ. This short-term activation of IDO1 occurs as part of the innate immune response and helps to inhibit the growth of pathogens and parasites. IDO1 activation can also promote host immuno tolerance by exerting an immunosuppressive effect. IDO1 expression by tumor cells plays a susbtancial role in tumor immune tolerance, aiding tumors in evading detection and destruction. Chronic induction of IDO1 expression has been found in cancer patients increased IDO1 activity is correlated with negative prognosis. IDO1 has thus become an attractice pharmacological target for development of novel antineoplastics and adjuvants to increase the efficacy of conventional chemotherapy.
-
Platform
Microplate reader
Properties
-
Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests IDO1 Assay Buffer 1 x 50ml Antioxidant Mix (100X) 1 vial N-formylkynurenine Standard 1 vial IDO1 Substrate (L-tryptophan) 1 vial IDO1 Inhibitor (IDO5L) 1 vial Fluorogenic Developer Solution 1 x 5ml Recombinant Human IDO1 1 vial Microplate Sealing Film 1 unit -
Research areas
-
Function
Catalyzes the cleavage of the pyrrol ring of tryptophan and incorporates both atoms of a molecule of oxygen. -
Pathway
Amino-acid degradation; L-tryptophan degradation via kynurenine pathway; L-kynurenine from L-tryptophan: step 1/2. -
Sequence similarities
Belongs to the indoleamine 2,3-dioxygenase family. - Information by UniProt
-
Alternative names
- 3-dioxygenase
- I23O1_HUMAN
- IDO
see all
Images
-
N-formylkynurenine (NFK) standard curve.
The reaction of 1 mole of NFK with Fluorogenic Developer corresponds to the metabolism of one mole of L-tryptophan by IDO1.
-
Measurement of IDO1 Positive Control in presence and absence of 1 µM of the included selective inhibitor IDO5L. -
IDO1 activity in lysates (30 µL) of human cancer cell lines stimulated with vehicle (dH2O) or 100 ng/mL human interferon-? for 24 hours prior to assay.
