RNA-DNA Hybrid anticorps

Détails pour le produit réf. ABIN4889499
Toutes les espèces
Clonalité (Clone)
Monoclonal ()
Chromatin Immunoprecipitation (ChIP), Immunofluorescence (IF), Immunohistochemistry (IHC), In situ hybridization (ISH)
'Independent Validation' signe
Antigène RNA-DNA Hybrid
Application validée Surface Plasmon Resonance
Contrôle positif RNA-DNA Hybrid complexes
Contrôle négative ssDNA, dsDNA

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

Anticorps primaire ABIN4889499
  • 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.

  • 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.

Validation Images
Surface Plasmon Resonance validation image for anti-RNA-DNA Hybrid antibody (ABIN4889499) The sensor response to the binding of RNA-DNA-hybrid antibody (left) and correspondin...
Immunogène DNA-RNA duplex (random RNA-DNA heteropolymer)
Clone D5H6
Isotype IgG
Specificité Reacts with RNA-DNA hybrid
Purification Purified (protein A)
Indications d'application

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

Restrictions For Research Use only
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