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Bevacizumab ELISA Kit

Reaktivität: Human, Maus, Ratte Colorimetric Sandwich ELISA 1.56-50 ng/mL Plasma, Serum
Produktnummer ABIN4886390
  • Target Alle Bevacizumab Produkte
    Bevacizumab
    Reaktivität
    Human, Maus, Ratte
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Detektionsbereich
    1.56-50 ng/mL
    Untere Nachweisgrenze
    1.56 ng/mL
    Applikation
    ELISA
    Verwendungszweck
    Quantification of Bevacizumab in biological matrices
    Proben
    Plasma, Serum
    Analytische Methode
    Quantitative
    Spezifität
    Bevacizumab (Avastin)
    Kreuzreaktivität (Details)
    hIgG1, rituximab, and infliximab prepared at 250 ng/mL were assayed and exhibited no crossreactivity or interference.
    Sensitivität
    1.5 ng/mL
    Bestandteile
    Coated microtiter plate, 96 wells
    Calibrator diluent. - 1.8ml
    Calibrator 12ul
    10X wash buffer - 25ml
    Assay buffer - 50ml
    1000X detection reagent - 17ul
    TMB - 12ml
    TMB stop solution - 12ml
    Plate sealers - 3
    Benötigtes Material
    Precision pipettes calibrated to deliver 5-1000μL
    Multi-channel pipette calibrated to deliver 50-200μL
    Plate shaker
    Disposable tips
    Vortex-Mixer
    Distilled or de-ionized water
    Microplate reader capable of reading 450nm with background subtrac
  • Applikationshinweise
    Optimal working dilution should be determined by the investigator.
    Probenmenge
    15 μL
    Testdauer
    2.5 h
    Plattentyp
    Pre-coated
    Protokoll
    The Bevacizumab ELISA kit is designed to measure free Bevacizumab with high specificity and sensitivity. This assay employs the sandwich enzyme immunoassay technique. A precoated anti-Bevacizumab 96 well plate is provided. Calibrator, quality control samples and test samples are pipetted into the appropriate wells. Bevacizumab present in biological matrices is bound by the immobilized capture antibody. After washing away any unbound substances, enzyme linked detection antibody is added to the wells. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of Bevaciuzumab present in test samples and the concentration is calculated from the standard series.
    Aufbereitung der Reagenzien

    Prepare only the appropriate amount of required reagent on the day of use. Store all reagents as per instructions stated on the label. 1. Wash Buffer (1X) Preparation: Dilute wash buffer concentrate with ultra-pure water 1/10 before use (for example add 20 mL concentrate to 180 mL ultra-pure water). Mix well. 2. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 11 μL concentrate to 11 mL of assay buffer). Mix well. 3. Preparation of Calibrators: Prepare calibrators with concentrations ranging from 2,500 ng/mL to 78 ng/mL. The following is an example calibrator curve.

    Probennahme
    This kit is compatible with EDTA-plasma, heparinplasma and serum samples. Samples can be stored at or below -20 °C for up to 1 year.
    Aufbereitung der Proben

    Dilute calibrators and test samples 1/50 with assay buffer (for example add 5μL of prepared calibrator or sample to 245μL of assay buffer). Mix well. Do not store diluted samples.

    Testdurchführung

    This assay employs the sandwich enzyme immunoassay technique. Anti- Bevacizumab is coated onto a 96 well microplate. Calibrator, quality control samples and test samples are pipetted into the appropriate wells. Bevacizumab present in biological matrices is bound by the immobilized capture antibody. After washing away any unbound substances, enzyme linked detection antibody is added to the wells. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of Bevacizumab present in test samples.

    Ergebnisberechnung
    1. Construct a standard curve by plotting the absorbance obtained from each standard against concentration. Use a 4 or 5 parameter curve fit. Alternatively a log-log curve fit may be used. 2. The concentration of the unknowns can be read directly from this standard curve using the absorbance value for each sample. 3. Any sample undiluted or diluted still reading greater than the highest standard should be diluted appropriately with calibrator diluent and retested. If the samples have been diluted, the concentration determined from the standard curve must be multiplied by the dilution factor.
    Testpräzision
    Precision: The precision was determined by analyzing samples prepared at 500 ng/mL in 6 replicates on 6 different occasions. Intra-assay coefficient of variation (CV) < 10%. Inter-assay CV < 10%.

    Recovery: 250 ng/mL of Bevacizumab was spiked in 10 lots of human serum. Recovery ranges are from 91-111% with an average recovery of 106%.
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Konservierungsmittel
    Without preservative
    Vorsichtsmaßnahmen
    Read manual completely before beginning
    Lagerung
    -20 °C
    Informationen zur Lagerung
    Store kit components at -20°C unless specified otherwise. DO NOT USE past kit expiration date. Some vials contain a small amount of reagents. Spin tubes on pulse setting prior to opening.
    Haltbarkeit
    12 months
  • Target Alle Bevacizumab Produkte
    Bevacizumab
    Abstract
    Bevacizumab Produkte
    Substanzklasse
    Antibody
    Hintergrund
    Bevacizumab (Trade name Avastin®) is a recombinant humanized monoclonal antibody that blocks angiogenesis by inhibiting vascular endothelial growth factor A (VEGF-A). VEGF-A stimulates angiogenesis in a variety of diseases, including cancer. Bevacizumab was the first clinically available angiogenesis inhibitor in the United States. Bevacizumab was approved by the U.S. Food and Drug Administration (FDA) for certain metastatic cancers. It received its first approval in 2004, for combined use with standard chemotherapy for metastatic colon cancer. It has since been approved for use in certain lung cancers, renal cancers, ovarian cancers, and glioblastoma multiforme of the brain. It had been approved for breast cancer, but that approval was withdrawn when later studies showed no evidence of effectiveness
    Gen-ID
    7422
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