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MAK356

Sigma-Aldrich

Indoleamine 2,3-Dioxygenase 1 (IDO1) Activity Assay Kit

sufficient for 100 fluorometric tests

Synonym(s):

IDO1 Activity Assay Kit

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About This Item

UNSPSC Code:
12352200

usage

sufficient for 100 fluorometric tests

detection method

fluorometric

relevant disease(s)

immunological diseases; cancer

General description

Indoleamine 2,3-Dioxygenase 1 (IDO1) is a cytoplasmic 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 immune tolerance by exerting an immunosuppressive effect. IDO1 expression by tumor cells play a substantial role in tumor immune tolerance, aiding tumors in evading detection and destruction. Chronic induction of IDO1 expression has been found in cancer patients and is correlated with negative prognosis. IDO1 has thus become an attractive pharmacological target for development of novel antineoplastics and adjuvants to increase the efficacy of conventional chemotherapy.

Features and Benefits

Compatible with high-throughput handling systems.

Suitability

Suitable for the assessment of native or recombinant IDO1 activity and the screening of drugs and novel ligands for induction or inhibition of cellular IDO1 activity in mammalian tissues expressing IDO1, cultured cells treated with IDO1-inducing cytokines (e.g. interferon-γ), recombinant IDO1 enzyme preparations, and IDO1-expressing heterologous cells.

Principle

The Indoleamine 2,3-Dioxygenase 1 Activity Assay Kit enables IDO1 activity to be easily determined in mammalian tissues and cell lines. The assay uses a fluorogenic developer that selectively reacts with NFK to produce a highly fluorescent product (ex = 402 nm/em = 488 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.

Pictograms

Corrosion

Signal Word

Warning

Hazard Statements

Precautionary Statements

Hazard Classifications

Met. Corr. 1

Storage Class Code

8A - Combustible corrosive hazardous materials

Flash Point(F)

188.6 °F

Flash Point(C)

87 °C


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Questions

  1. 1. What is the recommended concentration of protein to use per well?
    2. Do the IDO kit buffers contain inhibitors for endogenous enzymes that may metabolize N-formylkynurinine?
    3. Will anything in the kit be affected by using sonication instead of a dounce homogenizer?

    1 answer
    1. 1. There is no specific recommended amount of protein per well for testing. It is best to optimize this by testing a range of dilutions to determine the most effective concentration for the specific sample being used.
      2. The assay buffer does not contain any inhibitors.
      3. Mild sonication can be used instead of a dounce homogenizer, with low amplitude and short pulses, while keeping the samples on ice throughout the sonication process.

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