Multiplex Assay Kit for Alpha-Tocopherol (TCPa) ,etc. by FLIA (Flow Luminescence Immunoassay)

VE; Vitamin E; α-TCP; α-Tocopherol

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 94-101 98
EDTA plasma(n=5) 79-96 86
heparin plasma(n=5) 79-93 79

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alpha-Tocopherol (TCPa) ,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 Alpha-Tocopherol (TCPa) ,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 Alpha-Tocopherol (TCPa) ,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) 97-104% 87-105% 79-98% 81-97%
EDTA plasma(n=5) 97-104% 82-98% 81-104% 90-98%
heparin plasma(n=5) 88-97% 98-105% 95-104% 79-95%

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#:TCPa) 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 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
International Journal of Advanced Research Evaluation of enzymatic and non- enzymatic antioxidant status in seminal plasma of Iraqi Infertile Men Com:Source
Journal of Physiology and Pharmacology 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
Mycotoxin Res  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
The Journal of Nutritional Biochemistry Vitamin E and caloric restriction promote hepatic homeostasis through expression of connexin 26, N-cad, E-cad and cholesterol metabolism genes pubmed:27816814
Plant Physiol Biochem Ultrasonication of in vitro potato single node explants: Activation and recovery of antioxidant defence system and growth responses pubmed:29102903
PLoS One Oxidative stress in lung cancer patients is associated with altered serum markers of lipid metabolism Pubmed: 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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