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RNA-DNA Hybrid Antikörper

Dieses Anti--Antikörper ist ein Maus Monoklonal-Antikörper zur Detektion von in ChIP, IF, IHC und ISH. Geeignet für Alle Spezies. Unabhängig validiert für die Verwendung in Surface Plasmon Resonance.
Produktnummer ABIN4889499

Kurzübersicht für RNA-DNA Hybrid Antikörper (ABIN4889499)

Target

RNA-DNA Hybrid

Reaktivität

Alle Spezies

Wirt

Maus

Klonalität

Monoklonal

Applikation

Chromatin Immunoprecipitation (ChIP), Immunofluorescence (IF), Immunohistochemistry (IHC), In situ hybridization (ISH)

Klon

D5H6
  • Spezifität

    Reacts with RNA-DNA hybrid

    Aufreinigung

    Purified (protein A)

    Immunogen

    DNA-RNA duplex (random RNA-DNA heteropolymer)

    Isotyp

    IgG
  • Applikationshinweise

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

    Beschränkungen

    Nur für Forschungszwecke einsetzbar
  • Validierung #100000 (Surface Plasmon Resonance)
    'Independent Validation' Siegel
    by
    Institute of Photonics and Electronics AS CR, v.v.i.
    No.
    #100000
    Datum
    23.06.2015
    Antigen
    RNA-DNA Hybrid
    Chargennummer
    Validierte Anwendung
    Surface Plasmon Resonance
    Positivkontrolle
    RNA-DNA Hybrid complexes
    Negativkontrolle
    ssDNA, dsDNA
    Bewertung

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

    'Independent Validation' Siegel
    Validierungsbilder
    Protokoll
    Primärantikörper
    ABIN4889499
    Sekundärantikörper
    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.

    Anmerkungen
    • 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

    Rekonstitution

    Must be reconstituted in distilled water.

    Konzentration

    1 mg/mL

    Buffer

    Tris 0,1M, glycine 0,1M, sucrose 2 %

    Lagerung

    4 °C/-20 °C

    Informationen zur Lagerung

    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.

    Haltbarkeit

    24 months
  • Target

    RNA-DNA Hybrid

    Substanzklasse

    Nucleotide
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