Multiplex Assay Kit for Indoleamine-2,3-Dioxygenase (IDO) ,etc. by FLIA (Flow Luminescence Immunoassay)

CD107B; INDO; Indoleamine-Pyrrole 2,3 Dioxygenase

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Indoleamine-2,3-Dioxygenase (IDO) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Indoleamine-2,3-Dioxygenase (IDO) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 90-99 96
EDTA plasma(n=5) 92-101 96
heparin plasma(n=5) 95-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Indoleamine-2,3-Dioxygenase (IDO) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 89-98% 86-95% 80-99% 93-101%
EDTA plasma(n=5) 84-101% 89-96% 78-98% 82-93%
heparin plasma(n=5) 81-96% 92-101% 83-97% 80-101%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:IDO) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Blood CCL18 differentiates dendritic cells in tolerogenic cells able to prime regulatory T cells in healthy subjects Pubmed: 21803856
Clinical Transplantation Potential immunosuppressive function of plasma indoleamine 2,3-dioxygenase in patients with aGVHD after allo-HSCT Wiley: source
Indonesian Journal of Cancer Chemoprevention Secretion of Indoleamine 2, 3-Dioxygenase, an Immunomodulatory Substance, by Adipose-Derived Mesenchymal Stem Cell Iregway: Source
PLoS ONE CD40 Gene Silencing Reduces the Progression of Experimental Lupus Nephritis Modulating Local Milieu and Systemic Mechanisms PubMed: PMC3683035
Hepatology 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 carcinoma Pubmed: 23960017
Free Radical Biology and Medicine Indoleamine-2,3-dioxygenase elevated in tumor-initiating cells is suppressed by mitocans ScienceDirect: S0891584913006345
Frontiers in Oncology Cancer-Associated Fibroblasts from lung tumors maintain their immuno-suppressive abilities after high-dose irradiation Frontiersin:Source
Tissue Engineering Part A Immunogenicity and escape mechanisms of allogeneic tendon-derived stem cells Pubmed:24813640
Immunobiology. The expression of dendritic cell subsets in severe chronic rhinosinusitis with nasal polyps is altered Pubmed:24947894
Apoptosis Chronic hepatitis C virus infection triggers spontaneous differential expression of biosignatures associated with T cell exhaustion and apoptosis signaling in peripheral blood mononucleocytes Pubmed:25577277
Front Oncol Cancer-Associated Fibroblasts from Lung Tumors Maintain Their Immunosuppressive Abilities after High-Dose Irradiation PubMed: 26029659
Acta Universitatis Tamperensis Insights into the use of adipose stem cells for clinical cell therapy: Novel culturing conditions and characterization of multipotency and immunogenic properties Handle: 10024
PLoS One Respiratory Syncytial Virus-Infected Mesenchymal Stem Cells Regulate Immunity via Interferon Beta and Indoleamine-2, 3-Dioxygenase pubmed:27695127
Journal of Biomedical Nanotechnology Exploring Amino Acid-Capped Nanoparticles for Selective Anti-Parasitic Action and Improved Host Biocompatibility Pubmed:29883557
Biochemistry Mechanisms of Leptin and Ghrelin Action on Maturation and Functions of Dendritic Cells
American Journal of Pathology Cognitive Decline, Cerebral-Spleen Tryptophan Metabolism, Oxidative Stress, Cytokine Production, and Regulation of the Txnip Gene in 3xTg-AD Mice Pubmed: 30980800
international immunopharmacology Interaction of the immune-inflammatory and the kynurenine pathways in rats resistant to antidepressant treatment in model of depression Pubmed: 31176083
Stem Cell Reports Richard Schäfer,* Gabriele Spohn, Marco Bechtel, 2 Denisa Bojkova, 2 Patrick C. Baer, 3 Selim Kuçi, 4
International Journal of?Tryptophan?Research Altered Indoleamine 2, 3-Dioxygenase Production and Its Association to Inflammatory Cytokines in Peripheral Blood Mononuclear Cells Culture of Type 2 Diabetes?¡­ 33343199
Stem Cell Reports Richard Sch?fer,* Gabriele Spohn, Marco Bechtel, 2 Denisa Bojkova, 2 Patrick C. Baer, 3 Selim Ku?i, 4
Transl Psychiatry Associations between expression of indoleamine 2, 3-dioxygenase enzyme and inflammatory cytokines in patients with first-episode drug-naive … 34802039
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