RNA-DNA Hybrid 抗体

ABIN4889499 产品详细信息, 供应商: Log in to see
抗原
适用
所有种类
宿主
小鼠
克隆类型 (克隆位点)
单克隆 ()
应用范围
Chromatin Immunoprecipitation (ChIP), Immunofluorescence (IF), Immunohistochemistry (IHC), In situ hybridization (ISH)
选项
Supplier
Log in to see
Supplier Product No.
Log in to see
'独立验证'标志
抗原 RNA-DNA Hybrid
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.

Primary Antibody ABIN4889499
实验流程
  • 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.

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...
免疫原 DNA-RNA duplex (random RNA-DNA heteropolymer)
克隆位点 D5H6
亚型 IgG
特异性 Reacts with RNA-DNA hybrid
纯化方法 Purified (protein A)
应用备注

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

限制 仅限研究用
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
您还需要查找其他产品吗?