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RNA-DNA Hybrid anticorps

Reactivité: Toutes les espèces ChIP, IF, IHC, ISH Hôte: Souris Monoclonal D5H6 unconjugated
N° du produit ABIN4889499
  • Antigène
    RNA-DNA Hybrid
    Reactivité
    Toutes les espèces
    Hôte
    Souris
    Clonalité
    Monoclonal
    Application
    Chromatin Immunoprecipitation (ChIP), Immunofluorescence (IF), Immunohistochemistry (IHC), In situ hybridization (ISH)
    Specificité
    Reacts with RNA-DNA hybrid
    Purification
    Purified (protein A)
    Immunogène
    DNA-RNA duplex (random RNA-DNA heteropolymer)
    Clone
    D5H6
    Isotype
    IgG
  • Indications d'application

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

    Restrictions
    For Research Use only
  • Validation #100000 (Surface Plasmon Resonance)
    'Independent Validation' signe
    by
    Institute of Photonics and Electronics AS CR, v.v.i.
    No.
    #100000
    Date
    23.06.2015
    Antigène
    RNA-DNA Hybrid
    Numéro du lot
    Application validée
    Surface Plasmon Resonance
    Contrôle positif
    RNA-DNA Hybrid complexes
    Contrôle négative
    ssDNA, dsDNA
    Conclusion

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

    'Independent Validation' signe
    Validation Images
    Protocole
    Anticorps primaire
    ABIN4889499
    Anticorps secondaire
    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.

    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.

  • Format
    Lyophilized
    Reconstitution
    Must be reconstituted in distilled water.
    Concentration
    1 mg/mL
    Buffer
    Tris 0,1M, glycine 0,1M, sucrose 2 %
    Stock
    4 °C/-20 °C
    Stockage commentaire
    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.
    Date de péremption
    24 months
  • Antigène
    RNA-DNA Hybrid
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