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MMP1 ELISA Kit

MMP1 Reaktivität: Human Colorimetric Sandwich ELISA 8-18000 pg/mL Cell Culture Supernatant, Plasma, Serum
Produktnummer ABIN625053
  • Target Alle MMP1 ELISA Kits anzeigen
    MMP1 (Matrix Metallopeptidase 1 (Interstitial Collagenase) (MMP1))
    Reaktivität
    • 10
    • 4
    • 3
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    Human
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Detektionsbereich
    8-18000 pg/mL
    Untere Nachweisgrenze
    8 pg/mL
    Applikation
    ELISA
    Verwendungszweck
    Human MMP-1 ELISA Kit for cell culture supernatants, heparin treated plasma, and serum samples. EDTA and Citrate are not recommended.
    Proben
    Plasma, Cell Culture Supernatant, Serum
    Analytische Methode
    Quantitative
    Spezifität
    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.
    Sensitivität
    < 8 pg/mL
    Produktmerkmale
    • 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
    Bestandteile
    • 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
    Benötigtes Material
    • 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
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  • Applikationshinweise
    Recommended Dilution for serum and plasma samples2 - 10 fold
    Probenmenge
    100 μL
    Plattentyp
    Pre-coated
    Protokoll
    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.
    Aufbereitung der Reagenzien
    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.
    Testdurchführung
    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.
    Ergebnisberechnung

    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 %

    Testpräzision
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Handhabung
    Avoid repeated freeze-thaw cycles.
    Lagerung
    -20 °C
    Informationen zur Lagerung
    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.
    Haltbarkeit
    6 months
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    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." in: 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." in: 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." in: 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." in: 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." in: Environmental toxicology and pharmacology, Vol. 38, Issue 1, pp. 305-15, (2014) (PubMed).

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  • Target Alle MMP1 ELISA Kits anzeigen
    MMP1 (Matrix Metallopeptidase 1 (Interstitial Collagenase) (MMP1))
    Andere Bezeichnung
    MMP-1 (MMP1 Produkte)
    Synonyme
    CLG ELISA Kit, CLGN ELISA Kit, Mmp1a ELISA Kit, CG4859 ELISA Kit, Dm1-MMP ELISA Kit, Dmel\\CG4859 ELISA Kit, MMP-1 ELISA Kit, MMP1 ELISA Kit, Mmp 1 ELISA Kit, dMMP1 ELISA Kit, dm1-MMP ELISA Kit, dmmp1 ELISA Kit, l(2)k04809 ELISA Kit, mmp1 ELISA Kit, Mmp1 ELISA Kit, Mcol-A ELISA Kit, Mcola ELISA Kit, col4 ELISA Kit, mmp18 ELISA Kit, Clgn ELISA Kit, matrix metallopeptidase 1 ELISA Kit, Matrix metalloproteinase 1 ELISA Kit, matrix metalloproteinase 1 ELISA Kit, matrix metallopeptidase 1 (interstitial collagenase) ELISA Kit, matrix metallopeptidase 1a (interstitial collagenase) ELISA Kit, matrix metallopeptidase 1 S homeolog ELISA Kit, interstitial collagenase ELISA Kit, matrix metallopeptidase 8 L homeolog ELISA Kit, MMP1 ELISA Kit, Mmp1 ELISA Kit, RB11133 ELISA Kit, Mmp1a ELISA Kit, mmp1.S ELISA Kit, LOC100727966 ELISA Kit, mmp8.L ELISA Kit
    Hintergrund
    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%.
    Gen-ID
    4312
    UniProt
    P03956
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