Multiplex Assay Kit for Metallothionein 1 (MT1) ,etc. by FLIA (Flow Luminescence Immunoassay)

MT1A; MT1S; MTC

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 84-101 91
EDTA plasma(n=5) 86-94 90
heparin plasma(n=5) 79-98 84

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Metallothionein 1 (MT1) ,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 Metallothionein 1 (MT1) ,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 Metallothionein 1 (MT1) ,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) 80-105% 92-101% 91-98% 79-97%
EDTA plasma(n=5) 88-96% 83-99% 95-103% 87-101%
heparin plasma(n=5) 97-104% 90-101% 80-92% 98-105%

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#:MT1) 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
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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