CLIA Kit for Alpha-Tocopherol (TCPa) Pan-species (General) Competition CLIA

VE; Vitamin E; α-TCP; α-Tocopherol

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  • CLIA Kit for Alpha-Tocopherol (TCPa) Packages (Simulation)
  • CLIA Kit for Alpha-Tocopherol (TCPa) Packages (Simulation)
  • CLIA Kit for Alpha-Tocopherol (TCPa) Results demonstration
  • CCA922Ge.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of Alpha-Tocopherol (TCPa) and the recovery rates were calculated by comparing the measured value to the expected amount of Alpha-Tocopherol (TCPa) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-99 89
EDTA plasma(n=5) 80-92 85
heparin plasma(n=5) 93-104 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alpha-Tocopherol (TCPa) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Alpha-Tocopherol (TCPa) 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 Alpha-Tocopherol (TCPa) 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) 98-105% 96-104% 97-105% 88-95%
EDTA plasma(n=5) 80-102% 83-94% 80-98% 80-97%
heparin plasma(n=5) 81-98% 79-90% 91-99% 98-105%

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
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
Substrate A 1×10mL Substrate B 1×2mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 100µL Substrate Solution. Incubate 10 minutes at 37°C;
7. Read RLU value immediately.

CLIA Kit for Alpha-Tocopherol (TCPa)

Test principle

The microplate provided in this kit has been pre-coated with a monoclonal antibody specific to Alpha-Tocopherol (TCPa). A competitive inhibition reaction is launched between biotin labeled Alpha-Tocopherol (TCPa) and unlabeled Alpha-Tocopherol (TCPa) (Standards or samples) with the pre-coated antibody specific to Alpha-Tocopherol (TCPa). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Alpha-Tocopherol (TCPa) in the sample. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is reverse proportional to the Alpha-Tocopherol (TCPa) level in the sample or standard.

Citations

  • Evaluation of enzymatic and non- enzymatic antioxidant status in seminal plasma of Iraqi Infertile MenCom:Source
  • Effect of vitamin E on cerebral cortical oxidative stress and brain-derived neurotrophic factor gene expression induced by hypoxia and exercise in rats.PubMed: 25903950
  • The potential effects of antioxidant feed additives in mitigating the adverse effects of cornnaturally contaminated with Fusarium mycotoxins on antioxidant systems in the intestinal mucosa, plasma, and liver in weaned pigs.Pubmed:27021614
  • Vitamin E and caloric restriction promote hepatic homeostasis through expression of connexin 26, N-cad, E-cad and cholesterol metabolism genespubmed:27816814
  • Ultrasonication of in vitro potato single node explants: Activation and recovery of antioxidant defence system and growth responsespubmed:29102903
  • Oxidative stress in lung cancer patients is associated with altered serum markers of lipid metabolismPubmed: 30973911
  • Prooxidant effect of α-tocopherol in pancreatic tumors.
  • Vitamin A, C, D, E and B12 Levels in Leprosy: A Case Control Study

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