Multiplex Assay Kit for Metallothionein 2 (MT2) ,etc. by CBA (Cytometric Bead Array)

MT2A; CES1

Specificity

This assay has high sensitivity and excellent specificity for detection of Metallothionein 2 (MT2) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Metallothionein 2 (MT2) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Metallothionein 2 (MT2) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Metallothionein 2 (MT2) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 85-99 89
EDTA plasma(n=5) 87-95 91
heparin plasma(n=5) 85-95 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Metallothionein 2 (MT2) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Metallothionein 2 (MT2) ,etc. by CBA (Cytometric Bead Array) 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 2 (MT2) ,etc. by CBA (Cytometric Bead Array) 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% 93-101% 84-91% 94-103%
EDTA plasma(n=5) 90-99% 80-93% 81-96% 90-98%
heparin plasma(n=5) 80-89% 97-105% 78-102% 94-101%

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

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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Magazine Citations
Journal of Trace Elements in Medicine and Biology Metallothionein 2A gene polymorphism and trace elements in mother-newborn pairs in the Croatian population pubmed:29173474
Biochimica et Biophysica Acta GCN2 deficiency protects against high fat diet induced hepatic steatosis and insulin resistance in mice Pubmed:30006154
Arch Virol . Infectious bronchitis virus inhibits activation of the TLR7 pathway, but not the TLR3 pathway Pubmed: 32532134
Biomolecules Mercury Exposure Assessment in Mother–Infant Pairs from Continental and Coastal Croatia Pubmed: 32471193
Analytica Chimica Acta Gold nanoclusters as elemental label for the sequential quantification of apolipoprotein E and metallothionein 2A in individual human cells of the retinal … Pubmed:35361429
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