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Ferritin ELISA Kit

FE Reaktivität: Human Colorimetric Sandwich ELISA Cell Culture Supernatant, Plasma
Produktnummer ABIN612696
  • Target Alle Ferritin (FE) ELISA Kits anzeigen
    Ferritin (FE)
    Reaktivität
    • 5
    • 4
    • 3
    • 2
    • 2
    • 1
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    Human
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Untere Nachweisgrenze
    1.5 ng/mL
    Applikation
    ELISA
    Verwendungszweck
    The AssayMax Ferritin ELISA kit is designed for detection of human ferritin in plasma, serum, milk, and cell culture supernatants
    Marke
    AssayMax
    Proben
    Plasma, Cell Culture Supernatant
    Analytische Methode
    Quantitative
    Spezifität
    This assay recognizes both natural and recombinant human Ferritin.
    Bestandteile
    Ferritin Microplate: A 96 well polystyrene microplate (12 strips of 8 wells) coated with a polyclonal antibody against human Ferritin. Sealing Tapes: Each kit contains 3 pre-cut, pressure-sensitive sealing tapes that can be cut to fit the format of the individual assay. Ferritin Standard: Human Ferritin in a buffered protein base (100 ng, lyophilized). Biotinylated Ferritin Antibody (100x): A 100-fold biotinylated polyclonal antibody against Ferritin (80µl). EIA Diluent Concentrate (10x): A 10-fold concentrated buffered protein base (20 ml). Wash Buffer Concentrate (20x): A 20-fold concentrated buffered surfactant (30 ml, 2 bottles). Streptavidin-Peroxidase Conjugate (SP Conjugate): A 100-fold concentrate (80µl). 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
    < 4 h
    Plattentyp
    Pre-coated
    Protokoll
    This assay employs a quantitative sandwich enzyme immunoassay technique, which measures ferritin in less than 4 hours. A polyclonal antibody specific for ferritin has been pre-coated onto a microplate. Ferritin in standards and samples is sandwiched by the immobilized antibody and biotinylated polyclonal antibody specific for ferritin, 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 and bring all reagents to room temperature before use. If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved. EIA Diluent Concentrate (10x): Dilute the EIA Diluent Concentrate 1:10 with reagent grade water. Store for up to 1 month at 2-8°C. Ferritin Standard: Reconstitute the 100 ng of human Ferritin Standard with 2 ml of EIA Diluent to generate a stock solution of 50 ng/ml. Allow the standard to sit for 10 minutes with gentle agitation prior to making dilutions. Prepare duplicate or triplicate standard points by serially diluting the Standard solution (50 ng/ml) twofold with equal volume of EIA Diluent to produce 25, 12.5, 6.25, 3.125 and 1.563 ng/ml. Sample Diluent serves as the zero standard (0 ng/ml). Any remaining solution should be frozen at -20°C. Standard Point Dilution [Ferritin] (ng/ml) P1 1 part Standard (50 ng/ml) 50.00 P2 1 part P1 + 1 part EIA Diluent 25.00 P3 1 part P2 + 1 part EIA Diluent 12.50 P4 1 part P3 + 1 part EIA Diluent 6.250 P5 1 part P4 + 1 part EIA Diluent 3.125 P6 1 part P5 + 1 part EIA Diluent 1.563 P7 EIA Diluent 0.000 Biotinylated Ferritin Antibody (100x): Dilute the antibody 1:100 with EIA Diluent. Any remaining solution should be frozen at -20°C. Wash Buffer Concentrate (20x): Dilute the Wash Buffer Concentrate1: 20 with reagent grade water. SP Conjugate (100x): Dilute the conjugate 1:100 with EIA 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 1:10 into EIA Diluent. The undiluted samples can be stored at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles. 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. Dilute samples 1:10 into EIA Diluent. The undiluted samples can be stored at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles. Milk: Collect milk using sample tube. Centrifuge samples at 800 x g for 10 minutes and assay. Milk dilution is suggested at 1:4 into MIx Diluent, however, the user should determine the optimal dilution factor. 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. Dilute samples 1:10 into EIA Diluent. 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 with a sealing tape and incubate for two hours. Start the timer after the last sample addition. Wash five times with 200 µL of Wash Buffer manually. Invert the plate each time and decant the contents, hit it 4-5 times on absorbent paper towel to completely remove the liquid. If using a machine wash six times with 300 µL of Wash Buffer and then invert the plate, decant the contents, hit it 4-5 times on absorbent paper towel to completely remove the liquid. Add 50 µL of Biotinylated Ferritin Antibody to each well and incubate for one hour. Wash a microplate as described 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 a microplate as described above. Add 50 µL of Chromogen Substrate per well and incubate for about 10 minutes or till the optimal 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. If wavelength correction is available, subtract readings at 570 nm from those at 450 nm to correct optical imperfections. Otherwise, read the plate at 450 nm only. Please note that some unstable black particles may be generated at high concentration points after stopping the reaction for about 10 minutes, which will reduce the readings.

    Ergebnisberechnung

    Calculate the mean value of the duplicate or triplicate 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. 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 4.8 % and 7.2 % 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 EIA 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 zip-seal. May be stored for up to 1 month in a vacuum desiccator.
  • James, Friis, Woodd, Rehman, PrayGod, Kelly, Koethe, Filteau: "Minimal impact of an iron-fortified lipid-based nutrient supplement on Hb and iron status: a randomised controlled trial in malnourished HIV-positive African adults starting antiretroviral therapy." in: The British journal of nutrition, pp. 1-11, (2015) (PubMed).

    Horai, Koga, Fujikawa, Takatani, Nishino, Nakashima, Suzuki, Kawashiri, Iwamoto, Ichinose, Tamai, Nakamura, Ida, Kakugawa, Sakamoto, Ishimatsu, Mukae, Hamaguchi, Fujimoto, Kuwana, Origuchi, Kohno et al.: "Serum interferon-α is a useful biomarker in patients with anti-melanoma differentiation-associated gene 5 (MDA5) antibody-positive dermatomyositis. ..." in: Modern rheumatology / the Japan Rheumatism Association, Vol. 25, Issue 1, pp. 85-9, (2014) (PubMed).

    Alkhateeb, Zubritsky, Kinsman, Leitzel, Campbell-Baird, Ali, Connor, Lipton: "Elevation in multiple serum inflammatory biomarkers predicts survival of pancreatic cancer patients with inoperable disease." in: Journal of gastrointestinal cancer, Vol. 45, Issue 2, pp. 161-7, (2014) (PubMed).

    Alkhateeb, Leitzel, Ali, Campbell-Baird, Evans, Fuchs, Köstler, Lipton, Connor: "Elevation in inflammatory serum biomarkers predicts response to trastuzumab-containing therapy." in: PLoS ONE, Vol. 7, Issue 12, pp. e51379, (2013) (PubMed).

    Wang, Fu, Lien, Wang, Hsu, Wu, Chien, Cherng: "Effect of aerobic interval training on erythrocyte rheological and hemodynamic functions in heart failure patients with anemia." in: International journal of cardiology, (2012) (PubMed).

  • Target Alle Ferritin (FE) ELISA Kits anzeigen
    Ferritin (FE)
    Andere Bezeichnung
    Ferritin (FE Produkte)
    Synonyme
    1HCH ELISA Kit, BEST:LD36673 ELISA Kit, CG2216 ELISA Kit, Dmel\\CG2216 ELISA Kit, FER1 ELISA Kit, FER1HCH ELISA Kit, Fer-L ELISA Kit, Fer1 ELISA Kit, Fer1HC ELISA Kit, FerHCH ELISA Kit, HCH ELISA Kit, T21 ELISA Kit, T9 ELISA Kit, fer1 ELISA Kit, fer1hch ELISA Kit, l(3)00451 ELISA Kit, l(3)j10B4 ELISA Kit, SFerH-1 ELISA Kit, SOF-35 ELISA Kit, SOF-5L ELISA Kit, NV13011 ELISA Kit, ACP1 ELISA Kit, F8A5.24 ELISA Kit, F8A5_24 ELISA Kit, Ferritin 1 heavy chain homologue ELISA Kit, ferritin light chain ELISA Kit, ferritin ELISA Kit, chromosome 26 open reading frame, human C10orf131 ELISA Kit, hyaluronan and proteoglycan link protein 2 ELISA Kit, NAD(P)-linked oxidoreductase superfamily protein ELISA Kit, Fer1HCH ELISA Kit, H-1 ELISA Kit, YPDSF_1340 ELISA Kit, ftnA ELISA Kit, Ft ELISA Kit, C26H10orf131 ELISA Kit, HAPLN2 ELISA Kit, AT1G60730 ELISA Kit
    Hintergrund
    Ferritin is an iron storage protein. It consists of 24 subunits with combined molecular weight of 474,000. Serum ferritin level is related to body iron stores and is influenced by several diseases. High serum ferritin levels associate with iron overload [1], diabetes mellitus [2], Adult-onset Still disease (AOSD) [3], excessive macrophage activation [4], alcohol intake [5]. On the other hand, low level of ferritin is an indication of Iron Deficiency Anemia [6].
    Pathways
    Transition Metal Ion Homeostasis
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