ELISA Kit for Indoleamine-2,3-Dioxygenase (IDO) Rattus norvegicus (Rat) Sandwich ELISA

CD107B; INDO; Indoleamine-Pyrrole 2,3 Dioxygenase

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  • ELISA Kit for Indoleamine-2,3-Dioxygenase (IDO) Packages (Simulation)
  • ELISA Kit for Indoleamine-2,3-Dioxygenase (IDO) Packages (Simulation)
  • ELISA Kit for Indoleamine-2,3-Dioxygenase (IDO) Results demonstration
  • SEB547Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Indoleamine-2,3-Dioxygenase (IDO) and the recovery rates were calculated by comparing the measured value to the expected amount of Indoleamine-2,3-Dioxygenase (IDO) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-95 92
EDTA plasma(n=5) 97-104 101
heparin plasma(n=5) 96-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Indoleamine-2,3-Dioxygenase (IDO) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Indoleamine-2,3-Dioxygenase (IDO) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Indoleamine-2,3-Dioxygenase (IDO) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 82-102% 80-90% 81-96% 91-101%
EDTA plasma(n=5) 92-99% 94-103% 81-103% 83-99%
heparin plasma(n=5) 84-92% 82-95% 97-105% 95-102%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Indoleamine-2,3-Dioxygenase (IDO)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Indoleamine-2,3-Dioxygenase (IDO). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Indoleamine-2,3-Dioxygenase (IDO). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Indoleamine-2,3-Dioxygenase (IDO), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Indoleamine-2,3-Dioxygenase (IDO) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • CCL18 differentiates dendritic cells in tolerogenic cells able to prime regulatory T cells in healthy subjectsPubmed: 21803856
  • Potential immunosuppressive function of plasma indoleamine 2,3-dioxygenase in patients with aGVHD after allo-HSCTWiley: source
  • Secretion of Indoleamine 2, 3-Dioxygenase, an Immunomodulatory Substance, by Adipose-Derived Mesenchymal Stem CellIregway: Source
  • CD40 Gene Silencing Reduces the Progression of Experimental Lupus Nephritis Modulating Local Milieu and Systemic MechanismsPubMed: PMC3683035
  • Human CD14+ CTLA-4+ regulatory dendritic cells suppress T-cell response by cytotoxic T-lymphocyte antigen-4-dependent IL-10 and indoleamine-2,3-dioxygenase production in hepatocellular carcinomaPubmed: 23960017
  • Indoleamine-2,3-dioxygenase elevated in tumor-initiating cells is suppressed by mitocansScienceDirect: S0891584913006345
  • Cancer-Associated Fibroblasts from lung tumors maintain their immuno-suppressive abilities after high-dose irradiationFrontiersin:Source
  • Immunogenicity and escape mechanisms of allogeneic tendon-derived stem cellsPubmed:24813640
  • The expression of dendritic cell subsets in severe chronic rhinosinusitis with nasal polyps is alteredPubmed:24947894
  • Chronic hepatitis C virus infection triggers spontaneous differential expression of biosignatures associated with T cell exhaustion and apoptosis signaling in peripheral blood mononucleocytesPubmed:25577277
  • Cancer-Associated Fibroblasts from Lung Tumors Maintain Their Immunosuppressive Abilities after High-Dose IrradiationPubMed: 26029659
  • Insights into the use of adipose stem cells for clinical cell therapy: Novel culturing conditions and characterization of multipotency and immunogenic propertiesHandle: 10024
  • Respiratory Syncytial Virus-Infected Mesenchymal Stem Cells Regulate Immunity via Interferon Beta and Indoleamine-2, 3-Dioxygenasepubmed:27695127
  • Exploring Amino Acid-Capped Nanoparticles for Selective Anti-Parasitic Action and Improved Host BiocompatibilityPubmed:29883557
  • Mechanisms of Leptin and Ghrelin Action on Maturation and Functions of Dendritic Cells
  • Cognitive Decline, Cerebral-Spleen Tryptophan Metabolism, Oxidative Stress, Cytokine Production, and Regulation of the Txnip Gene in 3xTg-AD MicePubmed: 30980800
  • Interaction of the immune-inflammatory and the kynurenine pathways in rats resistant to antidepressant treatment in model of depressionPubmed: 31176083
  • Richard Schäfer,* Gabriele Spohn, Marco Bechtel, 2 Denisa Bojkova, 2 Patrick C. Baer, 3 Selim Kuçi, 4
  • Altered Indoleamine 2, 3-Dioxygenase Production and Its Association to Inflammatory Cytokines in Peripheral Blood Mononuclear Cells Culture of Type 2 Diabetes?¡­33343199
  • Richard Sch?fer,* Gabriele Spohn, Marco Bechtel, 2 Denisa Bojkova, 2 Patrick C. Baer, 3 Selim Ku?i, 4
  • Associations between expression of indoleamine 2, 3-dioxygenase enzyme and inflammatory cytokines in patients with first-episode drug-naive …34802039

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