ELISA Kit for Glutathione Peroxidase 1 (GPX1)

GSHPX1; Cellular glutathione peroxidase

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 83-97 92
EDTA plasma(n=5) 85-99 89
heparin plasma(n=5) 98-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glutathione Peroxidase 1 (GPX1) 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) 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) 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) 97-105% 87-101% 90-102% 80-91%
EDTA plasma(n=5) 78-89% 79-95% 86-102% 86-99%
heparin plasma(n=5) 83-93% 80-98% 82-104% 90-98%

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.

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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
AGE Consequences of age on ischemic wound healing in rats: altered antioxidant activity and delayed wound closure Pubmed:24443098
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
Cell Biol Int Influence of season on seminal antioxidant enzymes in Karan Fries bulls under tropical climatic conditions fileID:697741
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
Investigation of GPX1 Gene Expression, Serum GPX1 and Selenium Levels on Colorectal Cancer
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