ELISA Kit for Tissue Inhibitors Of Metalloproteinase 2 (TIMP2) Rattus norvegicus (Rat) Sandwich ELISA

CSC-21K; TIMP Metallopeptidase Inhibitor 2

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  • ELISA Kit for Tissue Inhibitors Of Metalloproteinase 2 (TIMP2) Packages (Simulation)
  • ELISA Kit for Tissue Inhibitors Of Metalloproteinase 2 (TIMP2) Packages (Simulation)
  • ELISA Kit for Tissue Inhibitors Of Metalloproteinase 2 (TIMP2) Results demonstration
  • SEA128Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Tissue Inhibitors Of Metalloproteinase 2 (TIMP2) and the recovery rates were calculated by comparing the measured value to the expected amount of Tissue Inhibitors Of Metalloproteinase 2 (TIMP2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 88-96 92
EDTA plasma(n=5) 86-98 91
heparin plasma(n=5) 94-102 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Tissue Inhibitors Of Metalloproteinase 2 (TIMP2) 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 2 (TIMP2) 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 2 (TIMP2) 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) 79-99% 93-104% 79-95% 85-92%
EDTA plasma(n=5) 87-96% 91-98% 96-104% 81-104%
heparin plasma(n=5) 82-95% 88-99% 98-105% 97-104%

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.

ELISA Kit for Tissue Inhibitors Of Metalloproteinase 2 (TIMP2)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Tissue Inhibitors Of Metalloproteinase 2 (TIMP2). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Tissue Inhibitors Of Metalloproteinase 2 (TIMP2). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Tissue Inhibitors Of Metalloproteinase 2 (TIMP2), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Tissue Inhibitors Of Metalloproteinase 2 (TIMP2) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Oral Administration of 3,3′-Diindolylmethane Inhibits Lung Metastasis of 4T1 Murine Mammary Carcinoma Cells in BALB/c MiceNutrition&Metabolism: source
  • Oral administration of benzyl-isothiocyanate inhibits solid tumor growth and lung metastasis of 4T1 murine mammary carcinoma cells in BALB/c miceSpringerLink: f3w831267267p277
  • Bone morphogenetic protein-2: a potential regulator in scleral remodelingPubMed: PMC2605409
  • Early outcome and blood-brain barrier integrity after co-administered thrombolysis and hyperbaric oxygenation in experimental strokePubMed: 21679435
  • 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 ApproachesPlosone: Source
  • Concentration of gelatinases and their tissue inhibitors in pancreatic inflammatory and neoplastic tumors and their influence on the early postoperative coursePubmed: 23585206
  • Analysis of the expression of nine secreted matrix metalloproteinases and their endogenous inhibitors in the brain of mice subjected to ischaemic strokePubmed:24671655
  • Impact of 6% hydroxyethyl starch 130/0.42 and 5% gelatin on renal function in a pediatric animal modelPubmed:24916275
  • The Complementary Effects of Atorvastatin and Exercise Treatment on the Composition and Stability of the Atherosclerotic Plaques in ApoE Knockout MicePubmed:25264981
  • Plasma inflammatory biomarkers for Huntington’s disease patients and mouse model Pubmed:25266150
  • Metalloproteinases and Their Tissue Inhibitors in Comparison between Different Chronic Pneumopathies in the HorseJournals: Mi
  • Plasma inflammatory biomarkers for Huntington’s disease patients and mouse modelPubMed: 25266150
  • The skin remodeling in type 1 diabetes and insulin resistance animal modelsPubMed: 26047379
  • The skin remodeling in type 1 diabetes and insulin resistance animal models.PubMed: 26047379
  • Assessment of MMP-1, MMP-8 and TIMP-2 in experimental periodontitis treated with kaempferolpmc:PMC4848383
  • From varices to venous ulceration: the story of chronic venous disease described by metalloproteinases.Pubmed:26991748
  • Metalloproteinases and their inhibitors are influenced by inhalative glucocorticoid therapy incombination with environmental dust reduction in equine recurrent airway obstruction.pubmed:27938355
  • Hypoxia enhances secretion of matrix metalloproteinases in bovine dermal fibroblasts:an in vitro approach to bovine digital dermatitis 10.108:09712119.2017.1335208
  • The MiR-495/Annexin A3/P53 Axis Inhibits the Invasion and EMT of Colorectal Cancer Cellspubmed:29224019
  • From varices to venous ulceration: the story of chronic venous disease described by metalloproteinasespubmed:26991748
  • Hypoxia enhances secretion of matrix metalloproteinases in bovine dermal fibroblasts: an approach to bovine digital dermatitis10.1080:09712119.2017.1335208
  • Metalloproteinases and their Inhibitors under the Course of Immunostimulation by CPG-ODN and Specific Antigen Inhalation in Equine Asthma
  • Correlation of Baseline Plasma and Inguinal Connective Tissue Metalloproteinases and Their Inhibitors With Late High-Pressure Endoleak After Endovascular …Pubmed: 31464166
  • Matrix metallopeptidase expression and modulation by transforming growth factor-β1 in equine endometrosisPubmed: 31980722
  • MMP-9 Concentration in Peritoneal Fluid Is a Valuable Biomarker Associated with Endotoxemia in Equine Colic33488296
  • Calcium Binding Protein S100A16 Expedites Proliferation, Invasion and EMT Process in Gastric Cancer34650982

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