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Trastuzumab Antibody ELISA 试剂盒

适用: 人, 小鼠, 大鼠 Colorimetric Bridging ELISA 31.25-125 ng/mL Plasma, Serum
产品编号 ABIN4886400
发货至: 中国
  • 抗原 See all Trastuzumab Antibody products
    Trastuzumab Antibody
    适用
    人, 小鼠, 大鼠
    检测方法
    Colorimetric
    实验类型
    Bridging ELISA
    检测范围
    31.25-125 ng/mL
    最低检测浓度
    31.25 ng/mL
    应用范围
    ELISA
    原理
    Quantification of antibodies to Trastuzumab
    样品类型
    Plasma, Serum
    Analytical Method
    Quantitative
    特异性
    Anti-Trastuzumab antibodies
    产品特性
    This immunogenicity assay employs the bridging ELISA technique.
    组件
    Coated microtiter plate, 96 wells
    QC samples - 4x50ul
    10X wash buffer - 25ml
    Assay buffer - 50ml
    1000X secondary antibody - 17ul
    1000X detection reagent - 17ul
    TMB - 12ml
    TMB stop solution - 12ml
    Plate sealers - 3
    试剂未包括
    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
  • 应用备注
    Optimal working dilution should be determined by the investigator.
    样本量
    15 μL
    实验时间
    3.5 h
    板类型
    Pre-coated
    实验流程
    The Trastuzumab immunogenicity assay employs the bridging ELISA technique. A precoated 96 well capture antibody plate is provided. Quality control and test samples are pipetted into the appropriate wells. Anti-Trastuzumab present in biological matrices binds the immobilized capture antibody. After washing away any unbound substances, secondary antibody is added to the wells and after a final wash a detection reagent is added. 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 anti-Trastuzumab present in test samples. Four levels of QC samples give a qualitative reference signal which can be used to determine the level (High, Medium, Low, Negative) of anti-Trastuzumab antibody in the unknown samples.
    试剂准备

    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 2 mL concentrate to 18 mL ultra-pure water). Mix well. 2. Secondary antibody (1X) Preparation: Dilute secondary antibody with assay buffer 1/1000 before use (for examples add 12 μL concentrate to 12 mL of assay buffer). Mix well. 3. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 12 μL concentrate to 12 mL of assay buffer). Mix well.

    样品收集
    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.
    样品制备

    Dilute QC samples and test samples 1/10 with assay buffer (for example add 30μL of prepared calibrator or sample to 270μL of assay buffer). Mix well. Do not store diluted samples. If test samples are out of range, then they may be further diluted.

    实验流程

    This immunogenicity assay employs the bridging ELISA technique. Capture antibody is precoated onto a 96 well microplate. Quality control and test samples are pipetted into the appropriate wells. AntiTrastuzumab present in biological matrices is bound by the immobilized capture antibody. After washing away any unbound substances, secondary antibody is added to the wells and after a final wash a detection reagent is added. 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 anti-Trastuzumab present in test samples. Three levels of QC samples give a qualitative reference signal which can be used to determine the level of anti-Trastuzumab antibody in the unknown samples. The color development is stopped and the intensity of the color is measured.

    结果分析
    1. Because anti-drug antibodies will vary in terms of affinity and concentration, this assay provides a qualitative readout. As such the user should use the comparable positive controls when comparing interassay results. The provided controls are tested for comparability between lots and can be traced. 2. The anti-drug antibody titers in the test samples will fall in the range of high, medium, low or negative. We recommend each lab develop their own statistical cutpoint using methodologies as described by G. Shankar, et al. (2008). (Recommendations for the validation of immunoassays used for detection of host antibodies against biotechnology products. J. Pharmaceutical and Biomedical Analysis 48:1267-1281). 3. Any sample undiluted or diluted still reading greater than the highest standard should be diluted appropriately with assay buffer and retested. If the samples have been diluted, the concentration determined from the standard curve must be multiplied by the dilution factor.
    实验精密度
    Intra-assay precision: < 10%
    Inter-assay precision: < 10%
    限制
    仅限研究用
  • 储存液
    Without preservative
    注意事项
    Read manual completely before beginning
    储存条件
    -20 °C
    储存方法
    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.
    有效期
    12 months
  • 抗原 See all Trastuzumab Antibody products
    Trastuzumab Antibody
    Abstract
    Trastuzumab Antibody 产品
    物质类
    Antibody
    背景
    Trastuzumab (Herceptin® ) is a humanized recombinant monoclonal antibody used for the treatment of primary breast cancers overexpressing human epidermal growth factor 2 (HER2). HER2 protein is overexpressed in 25-30 % of breast cancers.
    基因ID
    2064
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