MMP 9 Kit ELISA

N° du produit ABIN625060

Détail du produit MMP 9 Kit ELISA

Antigène: MMP 9 (MMP9) (Matrix Metallopeptidase 9 (Gelatinase B, 92kDa Gelatinase, 92kDa Type IV Collagenase) (MMP9))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 10-6000 pg/mL

Seuil minimal de détection: 10 pg/mL

Application: ELISA

Fonction: Human MMP-9 ELISA Kit for cell culture supernatants, heparin treated plasma, and serum samples. EDTA and Citrate are not recommended.

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 Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), MCP-1, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, MMP-1, - 2, -3, -10, PARC, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.

Sensibilité: < 10 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 samples300 - 3,000 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, 1x Assay Diluent (Item E) should be used for dilution of serum/plasma/culture supernatants/urine. Suggested dilution for normal serum/plasma: 300-3000 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 (Item E) should be diluted 5-fold with deionized or distilled water before use.
   4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL 1x Assay Diluent (Item E) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 80 µL MMP-9 standard from the vial of tem C, into a tube with 586.7 µL 1x Assay Diluent Buffer to prepare a 6000 pg/mL stock standard solution. Pipette 400myl 1x Assay Diluent into each tube. Use the stock standard solution to produce a Dilution series . Mix each tube thoroughly before the next transfer. 1x Assay Diluent serves as the zero standard (0 pg/mL). 200 µL 80 µL standard +586.7 µL 200myl 200 µL 200 µL 200 µL 200 µL 6000 2000 666.7 222.2 74.07 24.69 8.23 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 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 100-fold with 1x Assay Diluent and used in step 4 of Part VI Assay Procedure.
   7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 400-fold with 1x Assay Diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 30 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent to prepare a 400-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 Buffer Human MMP-9 concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of MMP-9 is typically less than 10 pg/mL.
Recovery: Recovery was determined by spiking various levels of human MMP-9 into normal human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 96.23 84-103 Plasma 94.64 83-102 Cell culture media 95.38 84-104
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 95 93 94 Range ( %) 84-103 83-102 85-104 1:4 Average % of Expected 96 97 96 Range ( %) 85-104 86-105 83-103
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 MMP 9 Kit ELISA (ABIN625060)

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).

Wu, Liu, Xie: "Osteopontin facilitates invasion in human trophoblastic cells via promoting matrix metalloproteinase-9 in vitro." dans: International journal of clinical and experimental pathology, Vol. 8, Issue 11, pp. 14121-30, (2016) (PubMed).

Devanarayanan, Nandeesha, Kattimani, Sarkar: "Relationship between matrix metalloproteinase-9 and oxidative stress in drug-free male schizophrenia: a case control study." dans: Clinical chemistry and laboratory medicine, Vol. 54, Issue 3, pp. 447-52, (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).

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).

Kapelouzou, Tsourelis, Kaklamanis, Degiannis, Kogerakis, Cokkinos: "Serum and tissue biomarkers in aortic stenosis." dans: Global cardiology science & practice, Vol. 2015, Issue 4, pp. 49, (2016) (PubMed).

Wang, Su, Yang, Qiao, Fang, Yu, Yang, Wang, Yin, Chen, Hong: "The influence of myeloid-derived suppressor cells on angiogenesis and tumor growth after cancer surgery." dans: International journal of cancer, Vol. 138, Issue 11, pp. 2688-99, (2016) (PubMed).

Naegelen, Plançon, Nicot, Kaoma, Muller, Vallar, Tschirhart, Bréchard: "An essential role of syntaxin 3 protein for granule exocytosis and secretion of IL-1?, IL-1?, IL-12b, and CCL4 from differentiated HL-60 cells." dans: Journal of leukocyte biology, Vol. 97, Issue 3, pp. 557-71, (2015) (PubMed).

Domienik-Kar?owicz, Rymarczyk, Dzikowska-Diduch, Lisik, Chmura, Demkow, Pruszczyk: "Emerging markers of atherosclerosis before and after bariatric surgery." dans: Obesity surgery, Vol. 25, Issue 3, pp. 486-93, (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).

Zhang, Lin, Jiang, Xu, Luo, Mo, Li, Chen: "Extensive serum biomarker analysis in patients with ST segment elevation myocardial infarction (STEMI)." dans: Cytokine, Vol. 76, Issue 2, pp. 356-62, (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).

Abdel-Latif: "Plasma Levels of Matrix Metalloproteinase (MMP)-2, MMP-9 and Tumor Necrosis Factor-α in Chronic Hepatitis C Virus Patients." dans: The open microbiology journal, Vol. 9, pp. 136-40, (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).

Gileles-Hillel, Alonso-Álvarez, Kheirandish-Gozal, Peris, Cordero-Guevara, Terán-Santos, Martinez, Jurado-Luque, Corral-Peñafiel, Duran-Cantolla, Gozal: "Inflammatory markers and obstructive sleep apnea in obese children: the NANOS study." dans: Mediators of inflammation, Vol. 2014, pp. 605280, (2014) (PubMed).

Khadjavi, Valente, Giribaldi, Prato: "Involvement of p38 MAPK in haemozoin-dependent MMP-9 enhancement in human monocytes." dans: Cell biochemistry and function, Vol. 32, Issue 1, pp. 5-15, (2014) (PubMed).

Kulkarni, Haldar, Nahire, Katti, Ambre, Muhonen, Shabb, Padi, Singh, Borowicz, Shrivastava, Katti, Reindl, Guo, Mallik: "Mmp-9 responsive PEG cleavable nanovesicles for efficient delivery of chemotherapeutics to pancreatic cancer." dans: Molecular pharmaceutics, Vol. 11, Issue 7, pp. 2390-9, (2014) (PubMed).

Détail du antigène

Antigène: MMP 9 (MMP9) (Matrix Metallopeptidase 9 (Gelatinase B, 92kDa Gelatinase, 92kDa Type IV Collagenase) (MMP9))

Autre désignation: MMP-9 (MMP9 Produits)

Synonymes: CLG4B Kit ELISA, GELB Kit ELISA, MANDP2 Kit ELISA, MMP-9 Kit ELISA, AW743869 Kit ELISA, B/MMP9 Kit ELISA, Clg4b Kit ELISA, pro-MMP-9 Kit ELISA, mmp-9 Kit ELISA, fgMMP-9 Kit ELISA, mmp9 Kit ELISA, ZFMMP-9 Kit ELISA, wu:fb02g06 Kit ELISA, wu:fb07b05 Kit ELISA, wu:fi98c09 Kit ELISA, wu:fj05a08 Kit ELISA, zgc:64165 Kit ELISA, clg4b Kit ELISA, gelb Kit ELISA, mandp2 Kit ELISA, matrix metallopeptidase 9 Kit ELISA, collagenase Kit ELISA, matrix metalloproteinase 9 Kit ELISA, matrix metallopeptidase 9 S homeolog Kit ELISA, MMP9 Kit ELISA, RB3913 Kit ELISA, BPSS0666 Kit ELISA, Sbal_3732 Kit ELISA, Bcer98_0486 Kit ELISA, Shew185_0630 Kit ELISA, Shal_3056 Kit ELISA, Sbal195_0657 Kit ELISA, Lbys_3550 Kit ELISA, Palpr_2084 Kit ELISA, Mmp9 Kit ELISA, mmp9 Kit ELISA, mmp9.S Kit ELISA

Sujet: Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that degrade extracellular matrix proteins. MMPs have been linked with a wide array of biological activities and play important roles during organ development and pathological processes. Collectively MMPs are key enzymes for the metabolism of extracellular matrix proteins, including fibrillar and non-fibrillar collagens, fibronectin, laminin and basement membrane or interstitial stroma glycoproteins. Under physiological conditions MMPs are involved in extracellular degradation and breakdown of matrix proteins during normal tissue remodelling processes such as wound healing, pregnancy, and angiogenesis. Human MMP-9 is a 92 kDa glycoprotein that plays a significant role in matrix remodeling, enzyme modulation, and cytokine/growth factor activation. MMP-9 is also known as gelatinase B based on its ability to degrade gelatin. The Human MMP-9 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human MMP-9 pro and active forms in serum, plasma (Collect plasma using heparin as an anticoagulant. EDTA and Citrate are not recommended), cell culture supernatants and urine. This assay employs an antibody specific for human MMP-9 coated on a 96-well plate. Standards and samples are pipetted into the wells and MMP-9 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human MMP-9 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 MMP-9 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: 4318

UniProt: P14780

Pathways: Cellular Response to Molecule of Bacterial Origin, Positive Regulation of Immune Effector Process, CXCR4-mediated Signaling Events