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

CD107B; INDO; Indoleamine-Pyrrole 2,3 Dioxygenase

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

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  • Multiplex Assay Kit for Indoleamine-2,3-Dioxygenase (IDO) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Indoleamine-2,3-Dioxygenase (IDO) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Indoleamine-2,3-Dioxygenase (IDO) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMB547Ra.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) ,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) 79-104 85
EDTA plasma(n=5) 97-105 101
heparin plasma(n=5) 78-98 90

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) 81-101% 94-102% 99-105% 96-105%
EDTA plasma(n=5) 80-99% 88-96% 93-104% 82-101%
heparin plasma(n=5) 80-89% 92-104% 82-99% 83-91%

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.

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

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

Giveaways

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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