Multiplex Assay Kit for Suppressors Of Cytokine Signaling 3 (SOCS3) ,etc. by FLIA (Flow Luminescence Immunoassay)

ATOD4; CIS3; Cish3; MGC71791; SOCS-3; SSI-3; SSI3; Cytokine-inducible SH2 protein 3; STAT-induced STAT inhibitor 3

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Suppressors Of Cytokine Signaling 3 (SOCS3) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Suppressors Of Cytokine Signaling 3 (SOCS3) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Suppressors Of Cytokine Signaling 3 (SOCS3) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Suppressors Of Cytokine Signaling 3 (SOCS3) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-104 96
EDTA plasma(n=5) 78-93 84
heparin plasma(n=5) 83-103 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Suppressors Of Cytokine Signaling 3 (SOCS3) ,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 Suppressors Of Cytokine Signaling 3 (SOCS3) ,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 Suppressors Of Cytokine Signaling 3 (SOCS3) ,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) 91-99% 86-97% 84-104% 86-102%
EDTA plasma(n=5) 93-103% 95-102% 93-101% 87-95%
heparin plasma(n=5) 78-91% 87-94% 98-105% 92-105%

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#:SOCS3) 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
The Journal of Biological Chemistry Human primary keratinocytes show a restricted ability to up-regulate suppressor of cytokine signaling (SOCS)3 compared to autologous macrophages Jcb: 280370
The Journal of Biological Chemistry Human Primary Keratinocytes Show Restricted Ability to Up-regulate Suppressor of Cytokine Signaling (SOCS)3 Protein Compared with Autologous Macrophages PubMed: PMC3323061
Journal of Reproductive Immunology What is the impact of Th1/Th2 ratio, SOCS3, IL17, and IL35 levels in unexplained infertility? Pubmed: 24368037
The Journal of Immunology Alveolar Epithelial Cell–Derived Prostaglandin E2 Serves as a Request Signal for Macrophage Secretion of Suppressor of Cytokine Signaling 3 during Innate Inflammation Pubmed:27183597
Scientific Reports The FXR Agonist, Obeticholic Acid, Suppresses HCC Proliferation & Metastasis: Role of IL-6/STAT3 Signalling Pathway pubmed:28970500
Journal of Immunological Methods Endogenous peroxidases in sputum interfere with horse-radish peroxidase-based ELISAs. pubmed:29154772
Turkish Journal Of Medical Sciences The impact of plasma SOCS3 levels and endometrial leukocytes on unexplained infertility Pubmed:29914245
Novel trajectories of Bromocriptine antidiabetic action: Leptin-IL-6/JAK2/p-STAT3/SOCS3, p-IR/p-AKT/GLUT4, PPARγ/Adiponectin, Nrf2/PARP-1, and GLP-1 articles:10.3389
JCI Insight Alveolar macrophage secretion of vesicular SOCS3 represents a platform for lung cancer therapeutics Pubmed: 31619584
European Journal of Pharmacology Olmesartan attenuates type 2 diabetes-associated liver injury: Cross-talk of AGE/RAGE/JNK, STAT3/SCOS3 and RAS signaling pathways Pubmed: 32067934
FASEB JOURNAL Resident alveolar macrophage‐derived vesicular SOCS3 dampens allergic airway inflammation Pubmed: 32030817
molecular immunology SOCS3-deficient lung epithelial cells uptaking neutrophil-derived SOCS3 worsens lung influenza infection Pubmed: 32645550
Experimental and Therapeutic Medicine Changes and clinical significance of serum inflammatory factors in the treatment of pregnancy hypertension syndrome with magnesium sulfate combined with … Pubmed: 32742411
NEUROCHEMICAL RESEARCH Modulation of the Nitric Oxide/BH4 Pathway Protects Against Irradiation-Induced Neuronal Damage 33755856
BIOMEDICINE & PHARMACOTHERAPY Potential contribution of ELISA and LFI Assays to assessment of the oxidative stress condition based on 8-oxodG biomarker 33957564
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