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RNA-DNA Hybrid 抗体

适用: 所有种类 ChIP, IF, IHC, ISH 宿主: 小鼠 Monoclonal D5H6 unconjugated
产品编号 ABIN4889499
发货至: 中国
  • 抗原
    RNA-DNA Hybrid
    适用
    所有种类
    宿主
    小鼠
    克隆类型
    单克隆
    应用范围
    Chromatin Immunoprecipitation (ChIP), Immunofluorescence (IF), Immunohistochemistry (IHC), In situ hybridization (ISH)
    特异性
    Reacts with RNA-DNA hybrid
    纯化方法
    Purified (protein A)
    免疫原
    DNA-RNA duplex (random RNA-DNA heteropolymer)
    克隆位点
    D5H6
    亚型
    IgG
  • 应用备注

    Working dilution: Optimal dilutions should be determined by the end user.

    限制
    仅限研究用
  • 生效 #100000 (Surface Plasmon Resonance)
    '独立验证'标志
    by
    Institute of Photonics and Electronics AS CR, v.v.i.
    No.
    #100000
    日期
    2015.06.23
    抗原
    RNA-DNA Hybrid
    Lot Number
    Method validated
    Surface Plasmon Resonance
    Positive Control
    RNA-DNA Hybrid complexes
    Negative Control
    ssDNA, dsDNA
    Notes

    The ability of the D5H6 antibody to recognize and bind RNA/DNA hetero-duplexes was approximately twice lower than expected.

    '独立验证'标志
    Validation Images
    Full Methods
    Primary Antibody
    ABIN4889499
    Secondary Antibody
    Full Protocol
    • For the experiments a four-channel spectroscopic SPR sensor with a temperature controller developed at IPE was used. In general, the SPR assay was based on the attachment of biotinylated DNA probes to the SPR sensor surface via the streptavidin-biotin interaction, with the streptavidin covalently attached to the alkanthiol self-assembled monolayer, subsequent forming of RNA/DNA hybrid duplexes and monitoring of their interaction with RNA-DNA-hybrid antibody.

    • The respective steps of the assay were as follows:

      1. Streptavidin was covalently attached to the sensor surface via amine coupling according to the previously published protocol (Vaisocherová et al. (2006) Biopolymers 82:394-398).
      2. Biotinylated DNA probes were immobilized in 10mM Tris buffer. The amount of probes was calculated to be of 1012 probes/cm2 and was kept the same across all experiments.
      3. Complementary miRNA strands were bound in 10mM Tris + 15mM MgCl2 to form RNA-DNA hybrid duplexes. The concentration range of miRNA used was 0.1-100nM.
      4. The D5H6 at a concentration of 1µg/ml was used in all experiments.
    • All the experiments were performed at the temperature of 25°C and the flow rate of 20µl/ml.

    Experimental Notes
    • The sensor response to the binding of D5H6 antibody obtained for various concentrations of miRNA and corresponding calibration curve are shown at Fig.1. The calibration curve is plotted as a dependence of the sensor response to the binding of D5H6 antibody on the concentration of miRNA and is linear up to the miRNA concentration of 5nM.

    • As a negative control, the ssDNA (only biotinylated DNA probe) or dsDNA (homo-duplex with DNA analogue of the miRNA) surface was used. In both cases, the nonspecific sensor response to the binding of D5H6 was below 2% which falls into the inter-assay variability range.

  • 状态
    Lyophilized
    溶解方式
    Must be reconstituted in distilled water.
    浓度
    1 mg/mL
    缓冲液
    Tris 0,1M, glycine 0,1M, sucrose 2 %
    储存条件
    4 °C/-20 °C
    储存方法
    Lyophilized powder stable for a minimum of 2 years at -20°C. Store reconstituted antibodies at +4°C. For extended periods store in aliquots at -20°C. Antibodies are guaranteed for 6 month from date of receipt.
    有效期
    24 months
  • 抗原
    RNA-DNA Hybrid
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