Telefon:
+49 (0)241 95 163 153
Fax:
+49 (0)241 95 163 155
E-Mail:
orders@antikoerper-online.de

IgA ELISA Kit

Reaktivität: Human Colorimetric Sandwich ELISA Cell Culture Supernatant, Milk, Plasma, Saliva, Serum, Urine
Produktnummer ABIN1440249
  • Target Alle IgA ELISA Kits anzeigen
    IgA
    Reaktivität
    • 9
    • 7
    • 7
    • 5
    • 5
    • 5
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 1
    • 1
    Human
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Untere Nachweisgrenze
    ~ 1.5 ng/mL
    Applikation
    ELISA
    Verwendungszweck
    The AssayMax Human IgA ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for detection of human IgA in plasma, serum, urine, saliva, milk, and cell culture supernatants. This assay employs a quantitative sandwich enzyme immunoassay technique that measures human IgA in less than 4 hours. A polyclonal antibody specific for human IgA has been pre-coated onto a 96-well microplate with removable strips. IgA in standards and samples is sandwiched by the immobilized antibody and the biotinylated polyclonal antibody specific for IgA, 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.
    Marke
    AssayMax
    Proben
    Serum, Milk, Saliva, Urine, Plasma, Cell Culture Supernatant
    Analytische Methode
    Quantitative
    Kreuzreaktivität (Details)
    Cross-Reactivity: Canine 25%, Monkey 5%, Mouse 1%, Swine 1%
    Produktmerkmale
    Standard Added Value: 5 - 50 ng/mL
    Bestandteile
    Human IgA Microplate: A 96-well polystyrene microplate (12 strips of 8 wells) coated with a polyclonal antibody against human IgA.
    Sealing Tapes: Each kit contains 3 precut, pressure sensitive sealing tapes that can be cut to fit the format of the individual assay.
    Human IgA Standard: Human IgA in a buffered protein base (200 ng, lyophilized).
    Biotinylated IgA Antibody (50x): A 50-fold concentrated biotinylated polyclonal antibody against IgA (140 µL).
    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, 2 bottles).
    Streptavidin-Peroxidase Conjugate (SP Conjugate): A 100-fold concentrate (80 µL).
    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, 200-1000 µL and multiple channel).
    Deionized or distilled reagent grade water.
    Featured
    Zu unserem meistverkauften IgA ELISA Kit
    Top Product
    Discover our top product IgA ELISA Kit
  • Probenmenge
    50 μL
    Testdauer
    < 4 h
    Plattentyp
    Pre-coated
    Protokoll
    Add 50 µL of standard/samples per well. Incubate 2 hours. Wash, then add 50 µL of biotinylated antibody per well. Incubate 1 hour. Wash, then add 50 µL of SP per well. Incubate 30 minutes. Wash, then add 50 µL of Chromogen Substrate per well. Incubate 12 minutes. Add 50 µL of Stop Solution per well. Read at 450 nm immediately.
    Aufbereitung der Reagenzien

    Freshly dilute all reagents and bring all reagents to room temperature before use.
    MIX Diluent Concentrate (10x): If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved. 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 200 ng of IgA Standard with 2 mL of MIX Diluent to generate a solution of 100 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 (100 ng/mL) 1:2 with MIX Diluent to produce 50, 25, 12.5, 6.25, 3.13, and 1.56 ng/mL solutions. MIX Diluent serves as the zero standard 4 (0 ng/mL). Any remaining solution should be frozen at -20°C and use within 30 days.
    Biotinylated IgA Antibody (50x): Spin down the antibody briefly and dilute the desired amount of the antibody 1:50 with MIX Diluent. Any remaining solution should be frozen at -20°C.
    Wash Buffer Concentrate (20x): If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved. 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.

    Aufbereitung der Proben

    Plasma: Collect plasma using one-tenth volume of 0.1 M sodium citrate as an anticoagulant. Centrifuge samples at 3000 x g for 10 minutes. Dilute samples 1:80000 into MIX Diluent and assay. If necessary, dilute samples within the range of 1:20000 to 1:200000. 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 an anticoagulant.)
    Serum: Samples should be collected into a serum separator tube. After clot formation, centrifuge samples at 3000 x g for 10 minutes and remove serum. Dilute samples 1:80000 into MIX Diluent and assay. If necessary, dilute samples within the range of 1:20000 to 1:200000. 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 3000 x g for 10 minutes to remove debris. Collect supernatants and assay. Store the remaining samples at -20°C or below. Avoid repeated freeze-thaw cycles.
    Urine: Collect urine using sample pot. Centrifuge samples at 800 x g for 10 minutes. Dilute urine 1:20 with MIX Diluent and assay. If necessary, dilute samples within the range of 1:10 to 1:100. Store samples at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles.
    Saliva: Collect saliva using sample tube. Centrifuge samples at 800 x g for 10 minutes. Dilute saliva 1:2000 with MIX Diluent. If necessary, dilute samples within the range of 1:1000 to 1:4000. Store samples 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. Dilute milk 1:10000 with MIX Diluent. If necessary, dilute samples within the range of 1:2000 to 1:20000. Store samples at -20°C or below for up to 3 months. 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 desiccants inside. Reseal the pouch securely to minimize exposure to water vapor and store in a vacuum desiccator.
    Add 50 µL of IgA Standard or sample per well. Cover wells with a sealing tape and incubate for 2 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 4-5 times on absorbent material to completely remove the liquid. If using a machine, wash six times with 300 µL of Wash Buffer and then invert the plate, decanting the contents, hit 4-5 times on absorbent material to completely remove the liquid.
    Add 50 µL of Biotinylated IgA Antibody to each well and incubate for 1 hour.
    Wash the microplate as described above.
    Add 50 µL of Streptavidin-Peroxidase Conjugate to each well and incubate for 30 minutes. Turn on the microplate reader and set up the program in advance.
    Wash the microplate as described above.
    Add 50 µL of Chromogen Substrate per well and incubate for about 12 minutes or till the optimal blue color density develops. Gently tap 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 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 four-parameter or log-log logistic curve-fit.
    Determine the unknown sample concentration from the Standard Curve and multiply the value by the dilution factor.

    Testpräzision
    Intra-assay and inter-assay coefficients of variation were 4.9% and 7.3% respectively.
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Handhabung
    Prepare all reagents (working diluent buffer, wash buffer, standards, biotinylated antibody, and SP conjugate) as instructed, prior to running the assay.
    Prepare all samples prior to running the assay. The dilution factors for the samples are suggested in this protocol. However, the user should determine the optimal dilution factor.
    Spin down the SP conjugate vial and the biotinylated antibody vial before opening and using contents.
    The kit should not be used beyond the expiration date.
    The Stop Solution is an acidic solution.
    Lagerung
    4 °C/-20 °C
    Informationen zur Lagerung
    Store components of the kit at 2-8°C or -20°C upon arrival up to the expiration date.
    Store SP Conjugate and biotinylated antibody at -20°C.
    Store Microplate, Diluent Concentrate (10x), Wash Buffer, Stop Solution, and Chromogen Substrate at 2-8°C.
    Unused microplate wells may be returned to the foil pouch with the desiccant packs and resealed. May be stored for up to 1 month in a vacuum desiccator.
    Diluent (1x) may be stored for up to 1 month at 2-8°C.
    Store standard at 2-8°C before reconstituting with diluent and at -20°C after reconstituting with diluent.
  • Target Alle IgA ELISA Kits anzeigen
    IgA
    Abstract
    IgA Produkte
    Synonyme
    MB-1 ELISA Kit, IG-alpha ELISA Kit, IGA ELISA Kit, IgA ELISA Kit, Igh-2 ELISA Kit, CD79a molecule ELISA Kit, immunoglobulin heavy constant alpha ELISA Kit, CD79A ELISA Kit, Igha ELISA Kit
    Substanzklasse
    Antibody
    Hintergrund
    Human Immunoglobulin A (IgA) is the most abundant antibody isotype in mucosal secretions and exists in two subclasses IgA-1 and IgA-2. While circulating serum IgA-1 occurs mainly in the monomeric 160 kDa form, mucosal secretary IgA-2 is in dimeric form and serves as the first line of defense against microorganisms through immune exclusion. Selective IgA deficiency is the most common primary immunodeficiency observed by a maturation defect in B cells to produce IgA. IgA nephropathy is the primary glomerulonephritis characterized by IgA deposition in the kidney and associated with a dysregulation of the immune response.
Sie sind hier:
Kundenservice