ELISA Kit for Metallothionein 1 (MT1)

MT1A; MT1S; MTC

Specificity

This assay has high sensitivity and excellent specificity for detection of Metallothionein 1 (MT1).
No significant cross-reactivity or interference between Metallothionein 1 (MT1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Metallothionein 1 (MT1) and the recovery rates were calculated by comparing the measured value to the expected amount of Metallothionein 1 (MT1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 89-96 93
EDTA plasma(n=5) 92-99 95
heparin plasma(n=5) 79-96 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Metallothionein 1 (MT1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Metallothionein 1 (MT1) 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 Metallothionein 1 (MT1) 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) 83-92% 82-95% 79-102% 91-99%
EDTA plasma(n=5) 94-101% 95-102% 99-105% 97-105%
heparin plasma(n=5) 87-97% 78-99% 84-104% 80-96%

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
Biotechology letters Modulated expression of genes associated with NO signal transduction contributes to the cholesterol-lowering effect of electro-acupuncture SpringerLink: c1v4234150886817
International Journal of Diabetes in Developing Countries Evaluation of serum metallothionein-1, selenium, zinc, and copper in Ghanaian type 2 diabetes mellitus patients Springer: Source
Experimental Oncology Peculiarities of antioxidant system and iron metabolism in organism during development of tumor resistance to cisplatin Pubmed:25265354
Journal of Neurochemistry Neuroprotective effects of levetiracetam target xCT in astrocytes in parkinsonian mice. Pubmed:26485447
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
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
European Journal of Immunology Metallothionein‐1 suppresses rheumatoid arthritis pathogenesis by shifting the Th17/Treg balance Pubmed:30055006
Int Immunopharmacol Metallothionein-1 is associated with osteoarthritis disease activity and suppresses proinflammatory cytokines production in synovial cells Pubmed: 31465913
Influence of cadmium on blood and hair trace elements and biochemical markers
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