Multiplex Assay Kit for Glutathione Peroxidase 1 (GPX1) ,etc. by FLIA (Flow Luminescence Immunoassay)

GSHPX1; Cellular glutathione peroxidase

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Glutathione Peroxidase 1 (GPX1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Glutathione Peroxidase 1 (GPX1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Glutathione Peroxidase 1 (GPX1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Glutathione Peroxidase 1 (GPX1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 87-101 97
EDTA plasma(n=5) 86-98 94
heparin plasma(n=5) 86-104 95
sodium citrate plasma(n=5) 87-101 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glutathione Peroxidase 1 (GPX1) ,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 Glutathione Peroxidase 1 (GPX1) ,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 Glutathione Peroxidase 1 (GPX1) ,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) 93-102% 89-96% 83-94% 90-101%
EDTA plasma(n=5) 80-103% 82-92% 93-105% 78-96%
heparin plasma(n=5) 80-105% 99-105% 86-94% 90-102%
sodium citrate plasma(n=5) 80-98% 82-97% 83-92% 86-94%

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#:GPX1) 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
PLoS One. P2×7?Receptor in the Kidneys of Diabetic Rats Submitted to Aerobic Training or to N-Acetylcysteine Supplementation Plos:Source
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Anticancer Research Determination of Gene Expression and Serum Levels of MnSOD and GPX1 in Colorectal Cancer Pubmed:25550558
Adv Med Sci Genetic polymorphisms (Pro197Leu of Gpx1, +35A/C of SOD1, PubMed: 26674569
Applied Biological Research Seasonal Variation in Glutathione Peroxidase in Seminal Plasma of Karan Fries (Tharparkar X Holstein Friesian) Bulls Under Tropical Climatic Conditions publication:301246301
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Toxicology Mechanisms and Methods Evaluation of hepatorenal impairments in Wistar rats coexposed to low-dose lead, cadmium and manganese: insights into oxidative stress mechanism pubmed:27599793
Journal of Environmental Research and Public Health Reduced Dietary Selenium Impairs Vascular Function by Increasing Oxidative Stress in Sprague-Dawley Rat Aortas. pubmed:28574428
Pharmacological Reports Assessing the serum concentration levels of NT-4/5, GPX-1, TNF-α, and l-arginine as biomediators of depression severity in first depressive episode patients with and without posttraumatic stress disorder. pubmed:28958613
CNS Spectrums Peripheral levels of superoxide dismutase and glutathione peroxidase in youths in ultra-high risk for psychosis: a pilot study pubmed:29248027
Environmental toxicology and pharmacology Dyslipdemia induced by chronic low dose co-exposure to lead, cadmium and manganese in rats: the role of oxidative stress pubmed:28654832
REVISTA DE CHIMIE Selenium- Essential Antioxidant Elemen the example of autoimune thyroiditis pdf:44%20PREDA%20C%207%2017.pdf
Biological Rhythm Research  Seasonal variation of mitochondria activity related and heat shock protein genes in spermatozoa of Karan Fries bulls in tropical climate 10.1080:09291016.2017.1361584
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