ELISA Kit for Thioredoxin (Trx) Rattus norvegicus (Rat) Sandwich ELISA

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

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  • ELISA Kit for Thioredoxin (Trx) Packages (Simulation)
  • ELISA Kit for Thioredoxin (Trx) Packages (Simulation)
  • ELISA Kit for Thioredoxin (Trx) Results demonstration
  • SEA702Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 85-92 88
EDTA plasma(n=5) 98-105 101
heparin plasma(n=5) 89-105 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Thioredoxin (Trx) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Thioredoxin (Trx) 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) 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) 91-103% 96-105% 80-90% 99-105%
EDTA plasma(n=5) 82-95% 81-95% 78-93% 98-105%
heparin plasma(n=5) 87-99% 95-103% 78-91% 88-97%

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 Thioredoxin (Trx)

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 Thioredoxin (Trx). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Thioredoxin (Trx). 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 Thioredoxin (Trx), 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 Thioredoxin (Trx) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • The roles of thioredoxin and thioredoxin-binding protein-2 in endometriosis OxfordJournals: source
  • Altered glutamate cysteine ligase activity in peripheral blood mononuclear cells from patients with systemic lupus erythematosusPubmed:Pmc4061188
  • Alteration in the intrafollicular thiol–redox system in infertile women with endometriosisPubmed:25376627
  • Thalassemia major patients using iron chelators showed a reduced plasma thioredoxin level and reduced thioredoxin reductase activity, despite elevated oxidative stressPubmed:25564095
  • 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
  • Relationship between aortic wall oxidative stress/proteolytic enzyme expression and intraluminal thrombus thickness indicates a novel pathomechanism in the …Pubmed: 30351992
  • Redox, immune and genetic biomarker system for personalized treatments in colorectal cancerDoi: 10.4251/wjgo.v11.i2.117
  • The Beneficial Effects of Lactobacillus plantarum PS128 on High-Intensity, Exercise-Induced Oxidative Stress, Inflammation, and Performance in TriathletesPubmed: 30736479
  • Nilotinib enhances β-islets integrity and secretory functions in a rat model of STZ-induced diabetes mellitusPubmed: 31351037
  • Damage-Associated Molecular Patterns and Th-Cell-Related Cytokines Released after Progressive EffortPubmed: 32210109
  • Thioredoxin 1 is upregulated in the bone and bone marrow following experimental myocardial infarction: evidence for a remote organ responsePubmed: 33161477
  • Examination of androgenetic alopecia with serum biomarkersPubmed: 32969583
  • Hepatoprotective Effect of Saccharomyces Cervisciae Cell Wall Extract against Thioacetamide-induced Liver Fibrosis in Rats34159266
  • Carvacrol hinders the progression of hepatic fibrosis via targeting autotaxin and thioredoxin in thioacetamide-induced liver fibrosis in rat34155936

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