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Multiplex Assay Kit for Tissue Inhibitors Of Metalloproteinase 3 (TIMP3) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 96-103 101
EDTA plasma(n=5) 80-92 88
heparin plasma(n=5) 94-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Tissue Inhibitors Of Metalloproteinase 3 (TIMP3) ,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 Tissue Inhibitors Of Metalloproteinase 3 (TIMP3) ,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 Tissue Inhibitors Of Metalloproteinase 3 (TIMP3) ,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) 83-102% 93-101% 96-103% 85-93%
EDTA plasma(n=5) 84-101% 89-96% 78-98% 82-93%
heparin plasma(n=5) 99-105% 90-98% 97-105% 87-102%

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#:TIMP3) 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
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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