ELISA Kit for Superoxide Dismutase 3, Extracellular (SOD3)

SOD-3; EC-SOD; Extracellular Superoxide Dismutase Cu-Zn

Specificity

This assay has high sensitivity and excellent specificity for detection of Superoxide Dismutase 3, Extracellular (SOD3).
No significant cross-reactivity or interference between Superoxide Dismutase 3, Extracellular (SOD3) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Superoxide Dismutase 3, Extracellular (SOD3) and the recovery rates were calculated by comparing the measured value to the expected amount of Superoxide Dismutase 3, Extracellular (SOD3) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 96-103 101
EDTA plasma(n=5) 93-105 99
heparin plasma(n=5) 82-101 87

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Superoxide Dismutase 3, Extracellular (SOD3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Superoxide Dismutase 3, Extracellular (SOD3) 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 Superoxide Dismutase 3, Extracellular (SOD3) 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% 89-97% 98-105% 79-97%
EDTA plasma(n=5) 94-102% 91-101% 82-90% 89-102%
heparin plasma(n=5) 83-93% 84-98% 78-101% 86-98%

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
Current Alzheimer Research Comparison of Extracellular and Intracellular Blood Compartments Highlights Redox Alterations in Alzheimer's and Mild Cognitive Impairment Patients. pubmed:27748187
Frontiers in Microbiology Burn Serum Increases Staphylococcus aureus Biofilm Formation via Oxidative Stress. pubmed:28702016
Nature Communications SOD3 improves the tumor response to chemotherapy by stabilizing endothelial HIF-2α Pubmed:29422508
BMC Neuroscience Neuroprotective dobutamine treatment upregulates superoxide dismutase 3, anti-oxidant and survival genes and attenuates genes mediating inflammation Pubmed:29523072
Diabetes-Metabolism Research and Reviews Myeloperoxidase, superoxide dismutase‐3, cardiometabolic risk factors, and distal sensorimotor polyneuropathy: The KORA F4/FF4 study Pubmed:29577557
Advances in Medical Sciences Selected elements of extracellular matrix of the skin in diabetes and insulin resistance Pubmed: 31146169
Clinical Kidney Journal Serum uromodulin is inversely associated with biomarkers of subclinical inflammation in the population-based KORA F4 study
PLoS One Association of circulating MR-proADM with all-cause and cardiovascular mortality in the general population: Results from the KORA F4 cohort study Pubmed:34990470
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