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ELISA Kit for Tissue Inhibitors Of Metalloproteinase 1 (TIMP1)

EPO; CLGI; EPA; HCI; Erythroid Potentiating Activity; Collagenase Inhibitor; Metalloproteinase inhibitor 1; Fibroblast collagenase inhibitor

Specificity

This assay has high sensitivity and excellent specificity for detection of Tissue Inhibitors Of Metalloproteinase 1 (TIMP1).
No significant cross-reactivity or interference between Tissue Inhibitors Of Metalloproteinase 1 (TIMP1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Tissue Inhibitors Of Metalloproteinase 1 (TIMP1) and the recovery rates were calculated by comparing the measured value to the expected amount of Tissue Inhibitors Of Metalloproteinase 1 (TIMP1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-98 94
EDTA plasma(n=5) 81-90 86
heparin plasma(n=5) 80-94 84

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Tissue Inhibitors Of Metalloproteinase 1 (TIMP1) 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 1 (TIMP1) 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 1 (TIMP1) 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) 83-101% 82-96% 81-99% 90-99%
EDTA plasma(n=5) 87-99% 95-103% 92-101% 91-101%
heparin plasma(n=5) 93-104% 94-101% 92-99% 80-103%

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
TMB Substrate 1×9mL Stop Solution 1×6mL
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 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Reference
Experimental & Translational Stroke Medicine Early outcome and blood-brain barrier integrity after co-administered thrombolysis and hyperbaric oxygenation in experimental stroke PubMed: 21679435
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
Polski Przeglad Chirurgiczny Concentration of gelatinases and their tissue inhibitors in pancreatic inflammatory and neoplastic tumors and their influence on the early postoperative course Pubmed: 23585206
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
The Scientific World Journal Pass-predicted hepatoprotective activity of Caesalpinia sappan in thioacetamide-induced liver fibrosis in rats Pubmed:24701154
PLOS ONE The Complementary Effects of Atorvastatin and Exercise Treatment on the Composition and Stability of the Atherosclerotic Plaques in ApoE Knockout Mice Pubmed:25264981
Mediators of Inflammation Metalloproteinases and Their Tissue Inhibitors in Comparison between Different Chronic Pneumopathies in the Horse Journals: Mi
PLOS Adiponectin Attenuates Lung Fibroblasts Activation and Pulmonary Fibrosis Induced by Paraquat PubMed: 25945502
Canadian Journal of Physiology and Pharmacology Modified Citrus Pectin stops progression of liver fibrosis by inhibiting galactin-3 and inducing apoptosis of stellate cells Doi: Abs
PLoS One Silencing of Paternally Expressed Gene 10 Inhibits Trophoblast Proliferation and Invasion PubMed: 26680220
Kyselov Влияние диуретиков с антиальдостероновым действием на состояние межклеточного матрикса и раннее постинфарктное ремоделирование левого … Article: View
Оригинальные исследования / Original researches матриксу та ранн Article: Viewfile
Basic Research CTGF upreCavia (Guinea pig )lation correlates with MMP-9 level in airway remodeling in a murine model of asthma Journal:27699
J Orthop Res. Tetrandrine suppresses articular inflammatory response by inhibiting pro-inflammatory factors via NF-κB inactivation. Pubmed:26748661
Int Wound J. From varices to venous ulceration: the story of chronic venous disease described by metalloproteinases. Pubmed:26991748
Am J Pathol.  Serum Osteopontin as a Novel Biomarker for Muscle Regeneration in Duchenne Muscular Dystrophy. Pubmed:26963343
Journal of Orthopaedic Research Tetrandrine suppresses articular inflammatory response by inhibiting pro‐inflammatory factors via NF‐κB inactivation pubmed:26748661
Canadian Journal of Veterinary Research Evaluation of hyaluronic acid, procollagen type III N-terminal peptide, and tissue inhibitor of matrix metalloproteinase-1 as serum markers of canine hepatic fibrosis pubmed:27733785
BMC Veterinary Research Metalloproteinases and their inhibitors are influenced by inhalative glucocorticoid therapy incombination with environmental dust reduction in equine recurrent airway obstruction. pubmed:27938355
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