Multiplex Assay Kit for Matrix Metalloproteinase 7 (MMP7) ,etc. by FLIA (Flow Luminescence Immunoassay)

MPSL1; MAT; PUMP-1; Matrilysin Uterine; Matrilysin; Matrin; Pump-1 Protease; Uterine Metalloproteinase

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Matrix Metalloproteinase 7 (MMP7) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Matrix Metalloproteinase 7 (MMP7) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Matrix Metalloproteinase 7 (MMP7) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Matrix Metalloproteinase 7 (MMP7) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 94-102 98
EDTA plasma(n=5) 78-103 82
heparin plasma(n=5) 94-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Matrix Metalloproteinase 7 (MMP7) ,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 Matrix Metalloproteinase 7 (MMP7) ,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 Matrix Metalloproteinase 7 (MMP7) ,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) 96-104% 98-105% 80-90% 81-102%
EDTA plasma(n=5) 84-91% 85-99% 80-105% 94-105%
heparin plasma(n=5) 81-101% 99-105% 83-103% 78-96%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:MMP7) 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
British Journal of Pharmacology A novel, orally active LPA1 receptor antagonist inhibits lung fibrosis in the mouse bleomycin model PubMed: 20649573
Neuroimmunomodulation Upregulated Expression of Matrix Metalloproteinases and Tissue Inhibitors of Matrix Metalloproteinases in BALB/c Mouse Brain Challenged with Japanese Encephalitis Virus Pubmed: 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
Thromb Haemost. Analysis of the expression of nine secreted matrix metalloproteinases and their endogenous inhibitors in the brain of mice subjected to ischaemic stroke Pubmed:24671655
Histol Histopathol Estrogen-deficient osteoporosis enhances the recruitment and activity of osteoclasts by breast cancer cells PubMed: 26254457
VirusDisease Circulating levels of matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases during Japanese encephalitis virus infection Pubmed:26925446
Am J Physiol Lung Cell Mol Physiol Systematic phenotyping and correlation of biomarkers with lung function and histology in lung fibrosis Pubmed:26993522
Cellular Physiology and Biochemistry The MiR-495/Annexin A3/P53 Axis Inhibits the Invasion and EMT of Colorectal Cancer Cells pubmed:29224019
s-space Thyroid disrupting effects and associated mechanisms of TCPP in GH3 cell line and zebrafish (Danio rerio) larva and adult 10371/137708
Science Translational Medicine Large-scale proteomics identifies MMP-7 as a sentinel of epithelial injury and of biliary atresia. pubmed:29167395
Hepatology Diagnostic Accuracy of Serum Matrix Metalloproteinase‐7 for Biliary Atresia Pubmed: 30153340
Journal of Pediatrics Quantification of Serum Matrix Metallopeptide 7 Levels May Assist in the Diagnosis and Predict the Outcome for Patients with Biliary Atresia Pubmed: 30853207
Pediatrics Serum MMP-7 in the Diagnosis of Biliary Atresia Pubmed: 31604829
Biomed Pharmacother Human umbilical cord blood mononuclear cells protect against renal tubulointerstitial fibrosis in cisplatin-treated rats Pubmed: 31710895
LABORATORY INVESTIGATION The synthetic toxin biliatresone causes biliary atresia in mice Pubmed: 32681026
diagnostics Interstitial Score and Concentrations of IL-4R¦Á, PAR-2, and MMP-7 in Bronchoalveolar Lavage Fluid Could Be Useful Markers for Distinguishing Idiopathic Interstitial?¡­ 33924683
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