TIMP1 Kit ELISA

N° du produit ABIN625098

Détail du produit TIMP1 Kit ELISA

Antigène: TIMP1 (TIMP Metallopeptidase Inhibitor 1 (TIMP1))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 2-1200 pg/mL

Seuil minimal de détection: 2 pg/mL

Application: ELISA

Fonction: Human TIMP-1 ELISA Kit for cell culture supernatants, plasma, and serum samples.

Type d'échantillon: Plasma, Cell Culture Supernatant, Serum

Analytical Method: Quantitative

Specificité: This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human ANG, CD23, Eotaxin, GCSF, GM-CSF, GRO-alpha, GRO-beta, GRO-gamma, I-309, IFN-gamma, IL-1 alpha, IL-1 beta, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p40), IL-12 (p70), IL-15, IL-16, IP-10, MCP-1, MCP-2, MCP-3, MCP-4, MCSF, MIG, MIP-1 alpha, MIP-1 beta, NAP-2, PDGF, PF-4, PARC, SCF, SDF-1 alpha, TNFbeta, TGFbeta1, TGFbeta2, TGFbeta3, VEGF.

Sensibilité: 40 pg/mL

Attributs du produit:

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Ingrédients:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Matériel non inclus:

• Distilled or deionized water
• Precision pipettes to deliver 2 μL to 1 μL volumes
• Adjustable 1-25 μL pipettes for reagent preparation
• 100 μL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

Information d'application

Indications d'application: Recommended Dilution for serum and plasma samples5 - 100 fold

Volume d'échantillon: 100 μL

Plaque: Pre-coated

Protocole:

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 μL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Préparation des réactifs:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
   2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants and urine. Suggested dilution for normal serum/plasma: 5-100 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
   3. Assay Diluent B should be diluted 5-fold with deionized or distilled water.
   4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium and urine) into Item C vial to prepare a 0.1 myg/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 120 µL TIMP-1 standard from the vial of Item C, into a tube with 546.7 µL Assay Diluent A or 1x Assay Diluent B to prepare a 18000 pg/mL stock standard solution. Pipette 400 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 200 µL 120 µL standard +546.7 µL 200myl 200 µL 200 µL 200 µL 200 µL 18000 6000 2000 666.7 222.2 74.07 24.69 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
   5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
   6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
   7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) before use. HRP-Streptavidin concentrate should be diluted 300-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 40 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent B to prepare a 300-fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). Mix well .

Procédure de l'essai:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
   2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
   3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
   4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
   5. Discard the solution. Repeat the wash as in step
   6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
   7. Discard the solution. Repeat the wash as in step
   8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
   9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.

Calcul des résultats:

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent A Human TIMP-1 concentration (pg/mL) O D =4 50 n m 0.01 0.1 1 10 10 100 1,000 10,000 100,000 Assay Diluent B Human TIMP-1 concentration (pg/mL) O D =4 50 n m 0.01 0.1 1 10 10 100 1,000 10,000 100,000
Sensitivity: The minimum detectable dose of TIMP-1 is typically less than 40 pg/mL.
Recovery: Recovery was determined by spiking various levels of human TIMP-1 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 92.43 81-102 Plasma 93.67 83-102 Cell culture media 94.49 84-104
Linearity: Sample Type Serum Plasma Cell culture media 1:2 Average % of Expected 94 93 94 Range ( %) 82-102 82-103 83-102 1:4 Average % of Expected 95 96 96 Range ( %) 84-103 85-104 84-104
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Précision du teste: Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %

Restrictions: For Research Use only

Stockage

Conseil sur la manipulation: Avoid repeated freeze-thaw cycles.

Stock: -20 °C

Stockage commentaire: The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Date de péremption: 6 months

Références pour TIMP1 Kit ELISA (ABIN625098)

Hanedan Onan, Baykan, Sezer, Narin, Mavili, Baykan, Uzum, Narin: "Evaluation of Cardiovascular Changes in Children with BAVs." dans: Pediatric cardiology, Vol. 37, Issue 3, pp. 472-81, (2017) (PubMed).

Wang, Song, Chen, Yuan, Xu, Zhang, Tan, Yang, Yu, Lv: "The Long-Term Influence of Tissue Inhibitor of Matrix Metalloproteinase-1 in Patients with Mild to Moderate Coronary Artery Lesions in a Chinese Population: A 7-Year Follow-Up Study." dans: Cardiology, Vol. 132, Issue 3, pp. 151-8, (2016) (PubMed).

Prato, Khadjavi, Magnetto, Gulino, Rolfo, Todros, Cavalli, Guiot: "Effects of oxygen tension and dextran-shelled/2H,3H-decafluoropentane-cored oxygen-loaded nanodroplets on secretion of gelatinases and their inhibitors in term human placenta." dans: Bioscience, biotechnology, and biochemistry, Vol. 80, Issue 3, pp. 466-72, (2016) (PubMed).

Tian, Gu, Zhao, Li, Lu, Li: "Urinary Emmprin, matrix metalloproteinase 9 and tissue inhibitor of metalloproteinase 1 as potential biomarkers in children with ureteropelvic junction narrowing on conservative treatment." dans: Nephrology (Carlton, Vic.), Vol. 20, Issue 3, pp. 194-200, (2015) (PubMed).

Koob, Lim, Zabek, Massee: "Cytokines in single layer amnion allografts compared to multilayer amnion/chorion allografts for wound healing." dans: Journal of biomedical materials research. Part B, Applied biomaterials, Vol. 103, Issue 5, pp. 1133-40, (2015) (PubMed).

Humbert, Fanian, Lihoreau, Jeudy, Elkhyat, Robin, Courderot-Masuyer, Tauzin, Lafforgue, Haftek: "Mécano-Stimulation™ of the skin improves sagging score and induces beneficial functional modification of the fibroblasts: clinical, biological, and histological evaluations." dans: Clinical interventions in aging, Vol. 10, pp. 387-403, (2015) (PubMed).

Gulino, Magnetto, Khadjavi, Panariti, Rivolta, Soster, Argenziano, Cavalli, Giribaldi, Guiot, Prato: "Oxygen-Loaded Nanodroplets Effectively Abrogate Hypoxia Dysregulating Effects on Secretion of MMP-9 and TIMP-1 by Human Monocytes." dans: Mediators of inflammation, Vol. 2015, pp. 964838, (2015) (PubMed).

Garratt, Sutanto, Ling, Looi, Iosifidis, Martinovich, Shaw, Kicic-Starcevich, Knight, Ranganathan, Stick, Kicic: "Matrix metalloproteinase activation by free neutrophil elastase contributes to bronchiectasis progression in early cystic fibrosis." dans: The European respiratory journal, Vol. 46, Issue 2, pp. 384-94, (2015) (PubMed).

Khadjavi, Magnetto, Panariti, Argenziano, Gulino, Rivolta, Cavalli, Giribaldi, Guiot, Prato: "Chitosan-shelled oxygen-loaded nanodroplets abrogate hypoxia dysregulation of human keratinocyte gelatinases and inhibitors: New insights for chronic wound healing." dans: Toxicology and applied pharmacology, Vol. 286, Issue 3, pp. 198-206, (2015) (PubMed).

Guan, Gao, Xu, Lin, Tang, Gu, Liu, Li, Chen, Zhong: "Sputum matrix metalloproteinase-8 and -9 and tissue inhibitor of metalloproteinase-1 in bronchiectasis: clinical correlates and prognostic implications." dans: Respirology (Carlton, Vic.), Vol. 20, Issue 7, pp. 1073-81, (2015) (PubMed).

Ciaffi, Cavassini, Genne, Delhumeau, Spycher Elbes, Hill, Wandeler, Fehr, Stoeckle, Schmid, Hirschel, Montecucco, Calmy: "Switch to etravirine for HIV-positive patients receiving statin treatment: a prospective study." dans: European journal of clinical investigation, Vol. 45, Issue 7, pp. 720-30, (2015) (PubMed).

Basilico, Magnetto, DAlessandro, Panariti, Rivolta, Genova, Khadjavi, Gulino, Argenziano, Soster, Cavalli, Giribaldi, Guiot, Prato: "Dextran-shelled oxygen-loaded nanodroplets reestablish a normoxia-like pro-angiogenic phenotype and behavior in hypoxic human dermal microvascular endothelium." dans: Toxicology and applied pharmacology, Vol. 288, Issue 3, pp. 330-8, (2015) (PubMed).

Markiewicz, Pytel, Mucha, Szymanek, Szaflik, Szaflik, Majsterek: "Altered Expression Levels of MMP1, MMP9, MMP12, TIMP1, and IL-1? as a Risk Factor for the Elevated IOP and Optic Nerve Head Damage in the Primary Open-Angle Glaucoma Patients." dans: BioMed research international, Vol. 2015, pp. 812503, (2015) (PubMed).

Fernández-Yunquera, Ripoll, Bañares, Puerto, Rincón, Yepes, Catalina, Salcedo: "Everolimus immunosuppression reduces the serum expression of fibrosis markers in liver transplant recipients." dans: World journal of transplantation, Vol. 4, Issue 2, pp. 133-40, (2014) (PubMed).

P Popat, Bhavsar, Popat: "Gingival crevicular fluid levels of Matrix Metalloproteinase-1 (MMP-1) and Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) in periodontal health and disease." dans: Singapore dental journal, Vol. 35, pp. 59-64, (2014) (PubMed).

Kiliç, Uçar, Özdemir, Colak, Bal, Ertu?rul: "Circulating matrix metalloproteinases and tissue inhibitors of metalloproteinases levels in pediatric patients with congenital heart disease: Relationship to cardiac functions." dans: Anadolu kardiyoloji dergisi : AKD = the Anatolian journal of cardiology, Vol. 14, Issue 6, pp. 531-41, (2014) (PubMed).

Gluba-Brzózka, Michalska-Kasiczak, Franczyk-Skóra, Nocu?, Banach, Rysz: "Markers of increased cardiovascular risk in patients with chronic kidney disease." dans: Lipids in health and disease, Vol. 13, pp. 135, (2014) (PubMed).

Rolin, Binda, Tissot, Viennet, Saas, Muret, Humbert: "In vitro study of the impact of mechanical tension on the dermal fibroblast phenotype in the context of skin wound healing." dans: Journal of biomechanics, Vol. 47, Issue 14, pp. 3555-61, (2014) (PubMed).

Ghodpage, Kolte, Kolte, Gupta: "Influence of phase I periodontal therapy on levels of matrix metalloproteinase 1 and tissue inhibitor of metalloproteinase 1." dans: The Saudi dental journal, Vol. 26, Issue 4, pp. 171-5, (2014) (PubMed).

Hampel, Klonisch, Sel, Schulze, Garreis, Seitmann, Zouboulis, Paulsen: "Insulin-like factor 3 promotes wound healing at the ocular surface." dans: Endocrinology, Vol. 154, Issue 6, pp. 2034-45, (2013) (PubMed).

Détail du antigène

Antigène: TIMP1 (TIMP Metallopeptidase Inhibitor 1 (TIMP1))

Autre désignation: TIMP-1 (TIMP1 Produits)

Synonymes: TIMP-1 Kit ELISA, TIMP1 Kit ELISA, DKFZp468A0912 Kit ELISA, CLGI Kit ELISA, EPA Kit ELISA, EPO Kit ELISA, HCI Kit ELISA, TIMP Kit ELISA, Timp Kit ELISA, Clgi Kit ELISA, TIMP metallopeptidase inhibitor 1 Kit ELISA, tissue inhibitor of metalloproteinase 1 Kit ELISA, TIMP1 Kit ELISA, Timp1 Kit ELISA

Sujet: The biological activities of the proteases are subject to a complex regulation also involving specific inhibitors, which are called TIMP (Tissue inhibitor of metalloproteinases). TIMP-1 is a protein of 28 kDa. The disruption of the TIMP-1gene in pluripotent embryonic stem cells can increase the invasive properties of these cells in an in vitro assay. The Human TIMP-1 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human TIMP-1 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human TIMP-1 coated on a 96-well plate. Standards and samples are pipetted into the wells and TIMP-1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human TIMP-1 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of TIMP-1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.

ID gène: 7076

UniProt: P01033