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IL-6 ELISA Kit

IL6 Reaktivität: Human Colorimetric Sandwich ELISA Cell Culture Supernatant, Plasma
Produktnummer ABIN612713
  • Target Alle IL-6 (IL6) ELISA Kits anzeigen
    IL-6 (IL6) (Interleukin 6 (IL6))
    Reaktivität
    • 16
    • 10
    • 9
    • 6
    • 6
    • 4
    • 4
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Untere Nachweisgrenze
    10 pg/mL
    Applikation
    ELISA
    Verwendungszweck
    The AssayMax Human IL-6 ELISA kit is designed for detection of IL-6 in human plasma, serum or cell culture supernatants
    Marke
    AssayMax
    Proben
    Plasma, Cell Culture Supernatant
    Analytische Methode
    Quantitative
    Spezifität
    This assay recognizes both natural and recombinant human IL-6.
    Bestandteile
    IL-6 Microplate: A 96 well polystyrene microplate (12 strips of 8 wells) coated with a murine monoclonal antibody against IL-6. Sealing Tapes: Each kit contains 3 pre-cut, pressure-sensitive sealing tapes that can be cut to fit the format of the individual assay. IL-6 Standard: Human IL-6 in a buffered protein base (0.75 ng, lyophilized). Biotinylated IL-6 Antibody (100x): A 100-fold concentrated biotinylated polyclonal antibody against IL-6 (80µl). MIx Diluent Concentrate (10x): A 10-fold concentrated buffered protein base (30 ml). Wash Buffer Concentrate (20x): A 20-fold concentrated buffered surfactant (30 ml). Streptavidin-Peroxidase Conjugate (SP Conjugate): A 100-fold concentrate (90µl). 1 Chromogen Substrate: A ready-to-use stabilized peroxidase chromogen substrate tetramethylbenzidine (8 ml). Stop Solution: A 0.5 N hydrochloric acid to stop the chromogen substrate reaction (12 ml).
    Benötigtes Material
    Microplate reader capable of measuring absorbance at 450 nm. Pipettes (1-20 µL, 20-200 µL, 200-1000µLand multiple channel). Deionized or distilled reagent grade water
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  • Probenmenge
    50 μL
    Testdauer
    5 h
    Plattentyp
    Pre-coated
    Protokoll
    This assay employs a quantitative sandwich enzyme immunoassay technique that measures IL-6 in 5 hours. A murine monoclonal antibody specific for human IL-6 has been pre-coated onto a microplate. IL-6 in standards and samples is sandwiched by the immobilized antibody and a biotinylated polyclonal antibody specific for human IL-6, which is recognized by a streptavidin-peroxidase conjugate. All unbound material is then washed away and a peroxidase enzyme substrate is added. The color development is stopped and the intensity of the color is measured.
    Aufbereitung der Reagenzien

    Freshly dilute all reagents at room temperature before use. If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved. MIx Diluent Concentrate (10x): Dilute the MIx Diluent Concentrate 1:10 with reagent grade water. Store for up to 1 month at 2-8°C. Standard Curve: Reconstitute the 0.75 ng of human IL-6 Standard with 0.75 ml of MIx Diluent to generate a solution of 1 ng/ml. Allow the standard to sit for 10 minutes with gentle agitation prior to making dilutions. Prepare triplicate standard points by serially diluting the IL-6 standard solution two-fold with equal volume of MIx Diluent to produce 0.5, 0.25, 0.125, 0.063, 0.031 and 0.016 ng/ml. MIx Diluent serves as the zero standard (0 ng/ml). Any remaining solution should be frozen at -20°C. Standard Point Dilution [IL-6] (ng/ml) P1 Standard (1 ng/ml) 1.000 P2 1 part P1 + 1 part MIx Diluent 0.500 P3 1 part P2 + 1 part MIx Diluent 0.250 P4 1 part P3 + 1 part MIx Diluent 0.125 P5 1 part P4 + 1 part MIx Diluent 0.063 P6 1 part P5 + 1 part MIx Diluent 0.031 P7 1 part P6 + 1 part MIx Diluent 0.016 P8 MIx Diluent 0.000 Biotinylated IL-6 Antibody (100x): Spin down the antibody briefly and dilute the desired amount of the antibody 1:100 with MIx Diluent. Any remaining solution should be frozen at -20°C. Wash Buffer Concentrate (20x): Dilute the Wash Buffer Concentrate 1:20 with reagent grade water. SP Conjugate (100x): Spin down the SP Conjugate briefly and dilute the desired amount of the conjugate 1:100 with MIx Diluent. Any remaining solution should be frozen at -20°C.

    Probennahme
    Plasma: Collect plasma using one-tenth volume of 0.1 M sodium citrate as an anticoagulant. Centrifuge samples at 2000 x g for 10 minutes and assay. Dilute samples 1:2 with MIx Diluent. The undiluted samples can be stored at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles. (EDTA or Heparin can also be used as anticoagulant.) Serum: Samples should be collected into a serum separator tube. After clot formation, centrifuge samples at 2000 x g for 10 minutes. Remove serum and assay. Store serum at -20°C or below. The undiluted samples can be stored at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles. Cell Culture Supernatants: Centrifuge cell culture media at 2000 x g for 10 minutes to remove debris. Collect supernatants and assay. Store samples at -20°C or below. Avoid repeated freeze-thaw cycles.
    Testdurchführung

    Prepare all reagents, working standards and samples as instructed. Bring all reagents to room temperature before use. The assay is performed at room temperature (20 - 30 °C). Remove excess microplate strips from the plate frame and return them immediately to the foil pouch with desiccant inside. Reseal the pouch securely to minimize exposure to water vapor and store in a vacuum desiccator. Add 50 µL of Standard or sample per well. Cover wells and incubate for two hours. Start the timer after the last sample addition. Wash five times with 200 µL of Wash Buffer. Invert the plate and decant the contents, and hit it 4-5 times on absorbent paper towel to complete remove liquid at each step. Add 50 µL of Biotinylated IL-6 Antibody to each well and incubate for two hours. Wash five times with 200 µL of Wash Buffer as above. Add 50 µL of Streptavidin-Peroxidase Conjugate per well and incubate for 30 minutes. Turn on the microplate reader and set up the program in advance. Wash five times with 200 µL of Wash Buffer as above. Add 50 µL of Chromogen Substrate per well and incubate for approximately 12 minutes or till the optimal blue color density develops. Gently tap the plate to ensure thorough mixing and break the bubbles in the well with pipette tip. Add 50 µL of Stop Solution to each well. The color will change from blue to yellow. Read the absorbance on a microplate reader at a wavelength of 450 nm immediately.

    Ergebnisberechnung

    Calculate the mean value of the duplicate or triplicate readings for each standard and sample. To generate a Standard Curve, plot the graph using the standard concentrations on the x-axis and the corresponding mean 450 nm absorbance on the y-axis. The best-fit line can be determined by regression analysis using log-log or four-parameter logistic curve-fit. Determine the unknown sample concentration from the Standard Curve and multiply the value by the dilution factor. 3 Standard Curve The curve is provided for illustration only. A standard curve should be generated each time the assay is performed.

    Testpräzision
    Intra-assay and inter-assay coefficients of variation were 5.1% and 7.0% respectively.
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Handhabung
    The kit should not be used beyond the expiration date.
    Lagerung
    4 °C/-20 °C
    Informationen zur Lagerung
    Store kit at 2-8°C or -20°C upon arrival up to the expiration date. Opened MIx Diluent may be stored for up to 1 month at 2-8°C. Store reconstituted reagents at -20°C or below. Opened unused strip wells may return to the foil pouch with the desiccant pack, reseal along with zip-seal. May be stored for up to 1 month in a vacuum desiccator.
  • Boga, Alkim, Koksal, Bayram, Ozguven, Ergun, Neijmann, Ozgon, Alkim: "Increased Plasma Levels of Asymmetric Dimethylarginine in Nonalcoholic Fatty Liver Disease: Relation With Insulin Resistance, Inflammation, and Liver Histology." in: Journal of investigative medicine : the official publication of the American Federation for Clinical Research, (2015) (PubMed).

    Bircan, Inal, Ozcelik, Koc, Demirag, Moray, Kemik: "LigaSure® versus Clamp Tie Technique for Thyroid Surgery; Decreased Operative Time versus Increased Inflammatory Effect: a prospective randomized study." in: European review for medical and pharmacological sciences, Vol. 18, Issue 14, pp. 1997-2005, (2014) (PubMed).

    Niyazoglu, Baykara, Koc, Aydoğdu, Onaran, Dellal, Tasan, Sultuybek: "Association of PARP-1, NF-κB, NF-κBIA and IL-6, IL-1β and TNF-α with Graves Disease and Graves Ophthalmopathy." in: Gene, Vol. 547, Issue 2, pp. 226-32, (2014) (PubMed).

    Twu, de Miguel, Lustig, Stevens, Vashisht, Wohlschlegel, Johnson: "Trichomonas vaginalis Exosomes Deliver Cargo to Host Cells and Mediate Host∶Parasite Interactions." in: PLoS pathogens, Vol. 9, Issue 7, pp. e1003482, (2013) (PubMed).

  • Target Alle IL-6 (IL6) ELISA Kits anzeigen
    IL-6 (IL6) (Interleukin 6 (IL6))
    Andere Bezeichnung
    Interleukin-6 (IL-6) (IL6 Produkte)
    Synonyme
    BSF2 ELISA Kit, HGF ELISA Kit, HSF ELISA Kit, IFNB2 ELISA Kit, IL-6 ELISA Kit, Il-6 ELISA Kit, ILg6 ELISA Kit, Ifnb2 ELISA Kit, il6 ELISA Kit, CHIL-6 ELISA Kit, interleukin 6 ELISA Kit, interleukin-6 ELISA Kit, IL6 ELISA Kit, Il6 ELISA Kit, il-6 ELISA Kit, IL-6 ELISA Kit
    Hintergrund
    Interleukin-6 (IL-6) is a cytokine of approximately 26 kDa that is synthesized by T-cells, macrophages, B-cells, fibroblasts, endothelial cells, and epithelial cells. IL-6 acts in both pro- inflammatory and anti-inflammatory ways. When released systemically it stimulates the liver to produce proteins, such as C-reactive protein and fibrin, that are responsible for the acute-phase response. Besides the systemic acute phase reaction, IL-6 is associated with several acute and chronic inflammatory diseases, including rheumatoid arthritis, acute pancreatitis, viral and bacterial meningitis, and Alzheimer's disease. However, IL-6 can also down regulate the inflammatory reaction by suppressing the pro-inflammatory cytokines IL-1 and TNF, and protect against lung damage and septic shock.
    Pathways
    TLR Signalweg, Hormone Transport, Negative Regulation of Hormone Secretion, Myometrial Relaxation and Contraction, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Regulation of Carbohydrate Metabolic Process, Autophagie, Cell RedoxHomeostasis, Cancer Immune Checkpoints, Inflammasome
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