MMP1 Kit ELISA

N° du produit ABIN625053

Détail du produit MMP1 Kit ELISA

Antigène: MMP1 (Matrix Metallopeptidase 1 (Interstitial Collagenase) (MMP1))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 8-18000 pg/mL

Seuil minimal de détection: 8 pg/mL

Application: ELISA

Fonction: Human MMP-1 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-2, - 3, -9, -10, PARC, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.

Sensibilité: < 8 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 samples2 - 10 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: 2-10 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 and then add 400 µL 1x Assay Diluent (Item E) into Item C vial to prepare a 0.1 myg/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 120 µL MMP-1 standard from the vial of item C, into a tube with 546.7 µL 1x Assay Diluent Buffer (for serum/plasma/cell culture medium/urine) to prepare a 18000 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 120 µL standard +546.7 µL 200myl 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 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 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 440-fold with 1x Assay Diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 25 µL of HRP-Streptavidin concentrate into a tube with 11 ml 1x Assay Diluent to prepare a final 440 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-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 MMP-1 is typically less than 8 pg/mL.
Recovery: Recovery was determined by spiking various levels of human MMP-1 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 97.78 88-106 Plasma 99.59 90-107 Cell culture media 98.66 90-106
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 98 99 98 Range ( %) 90-106 89-106 89-107 1:4 Average % of Expected 97 95 96 Range ( %) 90-106 89-107 90-107
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 MMP1 Kit ELISA (ABIN625053)

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).

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).

Tiraravesit, Yakaew, Rukchay, Luangbudnark, Viennet, Humbert, Viyoch: "Artocarpus altilis heartwood extract protects skin against UVB in vitro and in vivo." dans: Journal of ethnopharmacology, Vol. 175, pp. 153-62, (2015) (PubMed).

Abd-Allah, El-Shal, Shalaby, Pasha, Abou El-Saoud, Abdel Galil, Mahmoud: "Influence of Matrix metalloproteinase 1 and 3 genetic variations on susceptibility and severity of juvenile idiopathic arthritis." dans: IUBMB life, Vol. 67, Issue 12, pp. 934-42, (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).

Park, Kim, Kim, Oh: "Antioxidant effects of the sarsaparilla via scavenging of reactive oxygen species and induction of antioxidant enzymes in human dermal fibroblasts." dans: Environmental toxicology and pharmacology, Vol. 38, Issue 1, pp. 305-15, (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).

Gotoh, Nagano, Tsukahara, Murofushi, Morohoshi, Otsuka, Murakami-Murofushi: "Cyclic phosphatidic acid relieves osteoarthritis symptoms." dans: Molecular pain, Vol. 10, pp. 52, (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).

Soydinç, Sak, Evliyaoğlu, Evsen, Turgut, Ozler, Tay, Gül: "Maternal plasma prolidase, matrix metalloproteinases 1 and 13, and oxidative stress levels in pregnancies complicated by preterm premature rupture of the membranes and chorioamnionitis." dans: Journal of the Turkish German Gynecological Association, Vol. 13, Issue 3, pp. 172-7, (2014) (PubMed).

Angelini, Zacarias Fluck, Pedersen, Parra-Palau, Guiu, Bernadó Morales, Vicario, Luque-García, Navalpotro, Giralt, Canals, Gomis, Tabernero, Baselga, Villanueva, Arribas: "Constitutive HER2 signaling promotes breast cancer metastasis through cellular senescence." dans: Cancer research, Vol. 73, Issue 1, pp. 450-8, (2013) (PubMed).

Yuan, Zhao, Yuan, Che, Duan, Liu, Dong: "Association of endothelin-1 expression and cartilaginous endplate degeneration in humans." dans: PLoS ONE, Vol. 8, Issue 4, pp. e60062, (2013) (PubMed).

Denner, Eschricht, Lauck, Semaan, Schlaermann, Ryu, Akyüz: "Modulation of cytokine release and gene expression by the immunosuppressive domain of gp41 of HIV-1." dans: PLoS ONE, Vol. 8, Issue 1, pp. e55199, (2013) (PubMed).

Chen, Wong, Tam, Li, Lam: "Activation of human fibroblast-like synoviocytes by uric acid crystals in rheumatoid arthritis." dans: Cellular & molecular immunology, Vol. 8, Issue 6, pp. 469-78, (2012) (PubMed).

Kraus, Winter, Jepsen, Jäger, Meyer, Deschner: "Interactions of adiponectin and lipopolysaccharide from Porphyromonas gingivalis on human oral epithelial cells." dans: PLoS ONE, Vol. 7, Issue 2, pp. e30716, (2012) (PubMed).

Sundrani, Chavan-Gautam, Pisal, Mehendale, Joshi: "Matrix metalloproteinase-1 and -9 in human placenta during spontaneous vaginal delivery and caesarean sectioning in preterm pregnancy." dans: PLoS ONE, Vol. 7, Issue 1, pp. e29855, (2012) (PubMed).

Park, Jang, Oh: "Juglans mandshurica leaf extract protects skin fibroblasts from damage by regulating the oxidative defense system." dans: Biochemical and biophysical research communications, Vol. 421, Issue 2, pp. 343-8, (2012) (PubMed).

Abd-Allah, Shalaby, Pasha, El-Shal, Abou El-Saoud: "Variation of matrix metalloproteinase 1 and 3 haplotypes and their serum levels in patients with rheumatoid arthritis and osteoarthritis." dans: Genetic testing and molecular biomarkers, Vol. 16, Issue 1, pp. 15-20, (2012) (PubMed).

Jacob, Eisemon, Sheibani-Rad, Patel, Jacob, Choueka: "Matrix metalloproteinase levels as a marker for rotator cuff tears." dans: Orthopedics, Vol. 35, Issue 4, pp. e474-8, (2012) (PubMed).

Détail du antigène

Antigène: MMP1 (Matrix Metallopeptidase 1 (Interstitial Collagenase) (MMP1))

Autre désignation: MMP-1 (MMP1 Produits)

Synonymes: CLG Kit ELISA, CLGN Kit ELISA, Mmp1a Kit ELISA, CG4859 Kit ELISA, Dm1-MMP Kit ELISA, Dmel\\CG4859 Kit ELISA, MMP-1 Kit ELISA, MMP1 Kit ELISA, Mmp 1 Kit ELISA, dMMP1 Kit ELISA, dm1-MMP Kit ELISA, dmmp1 Kit ELISA, l(2)k04809 Kit ELISA, mmp1 Kit ELISA, Mmp1 Kit ELISA, Mcol-A Kit ELISA, Mcola Kit ELISA, col4 Kit ELISA, mmp18 Kit ELISA, Clgn Kit ELISA, matrix metallopeptidase 1 Kit ELISA, Matrix metalloproteinase 1 Kit ELISA, matrix metalloproteinase 1 Kit ELISA, matrix metallopeptidase 1 (interstitial collagenase) Kit ELISA, matrix metallopeptidase 1a (interstitial collagenase) Kit ELISA, matrix metallopeptidase 1 S homeolog Kit ELISA, interstitial collagenase Kit ELISA, matrix metallopeptidase 8 L homeolog Kit ELISA, MMP1 Kit ELISA, Mmp1 Kit ELISA, RB11133 Kit ELISA, Mmp1a Kit ELISA, mmp1.S Kit ELISA, LOC100727966 Kit ELISA, mmp8.L Kit ELISA

Sujet: The Human MMP-1 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human MMP-1 pro and active forms in serum, 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-1 coated on a 96-well plate. Standards and samples are pipetted into the wells and MMP-1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human MMP-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 MMP-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: 4312

UniProt: P03956