Multiplex Assay Kit for Thioredoxin (Trx) ,etc. by FLIA (Flow Luminescence Immunoassay)

TXN; ADF; SASP; TRDX; TRX1; ATL-derived factor; Surface-associated sulphydryl protein

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 79-91 85
EDTA plasma(n=5) 83-101 86
heparin plasma(n=5) 78-104 96
sodium citrate plasma(n=5) 93-102 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Thioredoxin (Trx) ,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 Thioredoxin (Trx) ,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 Thioredoxin (Trx) ,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) 95-105% 97-105% 83-99% 94-105%
EDTA plasma(n=5) 84-96% 85-101% 97-104% 91-99%
heparin plasma(n=5) 95-102% 98-105% 80-89% 87-95%
sodium citrate plasma(n=5) 93-101% 92-101% 80-95% 94-105%

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#:Trx) 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
Human Reproduction The roles of thioredoxin and thioredoxin-binding protein-2 in endometriosis OxfordJournals: source
Exp Ther Med. Altered glutamate cysteine ligase activity in peripheral blood mononuclear cells from patients with systemic lupus erythematosus Pubmed:Pmc4061188
Reproduction Alteration in the intrafollicular thiol–redox system in infertile women with endometriosis Pubmed:25376627
Free Radical Research Thalassemia major patients using iron chelators showed a reduced plasma thioredoxin level and reduced thioredoxin reductase activity, despite elevated oxidative stress Pubmed:25564095
Antioxidants Comparison of the Pulmonary Oxidative Stress Caused by Intratracheal Instillation and Inhalation of NiO Nanoparticles when Equivalent Amounts of NiO Are Retained in the Lung. Pubmed:26797643
FASEB Journal Relationship between aortic wall oxidative stress/proteolytic enzyme expression and intraluminal thrombus thickness indicates a novel pathomechanism in the … Pubmed: 30351992
World Journal of Gastrointestinal Oncology Redox, immune and genetic biomarker system for personalized treatments in colorectal cancer Doi: 10.4251/wjgo.v11.i2.117
Nutrients The Beneficial Effects of Lactobacillus plantarum PS128 on High-Intensity, Exercise-Induced Oxidative Stress, Inflammation, and Performance in Triathletes Pubmed: 30736479
European Journal of Pharmacology Nilotinib enhances β-islets integrity and secretory functions in a rat model of STZ-induced diabetes mellitus Pubmed: 31351037
Journal of Clinical Medicine Damage-Associated Molecular Patterns and Th-Cell-Related Cytokines Released after Progressive Effort Pubmed: 32210109
HISTOCHEMISTRY AND CELL BIOLOGY Thioredoxin 1 is upregulated in the bone and bone marrow following experimental myocardial infarction: evidence for a remote organ response Pubmed: 33161477
Journal of Cosmetic Dermatology Examination of androgenetic alopecia with serum biomarkers Pubmed: 32969583
Heliyon Hepatoprotective Effect of Saccharomyces Cervisciae Cell Wall Extract against Thioacetamide-induced Liver Fibrosis in Rats 34159266
Hum Exp Toxicol Carvacrol hinders the progression of hepatic fibrosis via targeting autotaxin and thioredoxin in thioacetamide-induced liver fibrosis in rat 34155936
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