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CLIA Kit for Tissue Inhibitors Of Metalloproteinase 3 (TIMP3)

SFD; TIMP Metallopeptidase Inhibitor 3; Metallopeptidase Inhibitor 3; Sorsby Fundus Dystrophy; Pseudoinflammatory

Specificity

This assay has high sensitivity and excellent specificity for detection of Tissue Inhibitors Of Metalloproteinase 3 (TIMP3).
No significant cross-reactivity or interference between Tissue Inhibitors Of Metalloproteinase 3 (TIMP3) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Tissue Inhibitors Of Metalloproteinase 3 (TIMP3) and the recovery rates were calculated by comparing the measured value to the expected amount of Tissue Inhibitors Of Metalloproteinase 3 (TIMP3) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 96-104 99
EDTA plasma(n=5) 78-94 81
heparin plasma(n=5) 86-101 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Tissue Inhibitors Of Metalloproteinase 3 (TIMP3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Tissue Inhibitors Of Metalloproteinase 3 (TIMP3) 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 Tissue Inhibitors Of Metalloproteinase 3 (TIMP3) 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) 79-104% 91-105% 79-103% 94-102%
EDTA plasma(n=5) 83-91% 99-105% 82-97% 94-102%
heparin plasma(n=5) 99-105% 98-105% 84-99% 80-91%

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
Substrate A 1×10mL Substrate B 1×2mL
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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

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Magazine Citations
Neuroimmunomodulation Upregulated Expression of Matrix Metalloproteinases and Tissue Inhibitors of Matrix Metalloproteinases in BALB/c Mouse Brain Challenged with Japanese Encephalitis Virus Pubmed: source
Experimental Biology and Medicine Acidic pH conditions mimicking degenerative intervertebral discs impair the survival and biological behavior of human adipose-derived mesenchymal stem cells Rsmjournals: Source
PLoS ONE A Combinatorial Relative Mass Value Evaluation of Endogenous Bioactive Proteins in Three-Dimensional Cultured Nucleus Pulposus Cells of Herniated Intervertebral Discs: Identification of Potential Target Proteins for Gene Therapeutic Approaches Plosone: Source
Physiol. Res. The unwounded skin remodeling in animal models of diabetes types 1 and 2. Pubmed: 24020818
VirusDisease Circulating levels of matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases during Japanese encephalitis virus infection Pubmed:26925446
Acta Pharmacol Sin. Pharmacological evidence: a new therapeutic approach to the treatment of chronic heart failure through SUR2B/Kir6.1 channel in endothelial cells. pubmed:27890915
Biochemical genetics Comparison of Selected Protein Levels in Tumour and Surgical Margin in a Group of Patients with Oral Cavity Cancer pubmed:28421310
International Journal of Cancer Circulating and tissue IMP3 levels are correlated with poor survival in renal cell carcinoma Pubmed: 30650187
J. Pers. Med Altered TIMP-3 Levels in the Cerebrospinal Fluid and Plasma of Patients with Alzheimer's Disease Pubmed:35629249
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