Recombinant
RabMAb

Recombinant Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] (ab208198)

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Overview

  • Product name

    Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44]
    See all N6, N6-dimethyladenosine (m6,6A) primary antibodies

  • Description

    Rabbit monoclonal [EPR- 19831-44] to N6, N6-dimethyladenosine (m6,6A)

  • Host species

    Rabbit

  • Specificity

    Has been developed to discriminate between the modified base N6, N6-dimethyladenosine (m6,6A) and the unmodified counterpart Adenosine (A).

  • Tested applications

    Suitable for: IP, ELISAmore details

  • Species reactivity

    Reacts with: Synthetic fragment

  • Immunogen

    Chemical/ Small Molecule corresponding to N6, N6-dimethyladenosine (m6,6A).

  • Positive control

    • IP: 5' Biotin-mN.mN.mN.mN.mN.[m6,6A]*.mN.mN.mN.mN.mN 3' * - phosphorothioate protection. ELISA: Biotinylated m6,6A-modified oligonucleotides.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR- 19831-44

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab208198 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IP Use at an assay dependent concentration.

0.2 µg of ab208198 was coated onto 50 µL Dynabeads® sheep-anti-rabbit IgG.
ELISA Use a concentration of 0.005 - 4 µg/ml.

Target

  • Relevance

    N6,N6-dimethyladenosine(m6,6A) is an RNA modification found in rRNA in archaea, bacteria and eukaryotes.

  • Alternative names

    • Di-m6A antibody
    • N6, N6 dimethyladenosine antibody

Images

  • Immunoprecipitation - Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] (ab208198)
    Immunoprecipitation - Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] (ab208198)

    The IP was performed in a U-bottom non-adsorbing propylene 96-well plate.

    ab208198 (0.2 μg) was coated into Dynabeads® sheep-anti-rabbit IgG (50 μl) for 1h at RT.

    Unmodified/modified oligonucleotides (5 μM) were added to samples containing the antibody/bead complexes and incubated with agitation for 1 hour at RT.

    After washing, Peroxidase-conjugated Streptavidin was incubated at 1/1000 dilution with agitation for 1 hour at RT.

    ECL substrate was then added and the results read in a non-transparent 96-well plate with a digital detector and analyzed using ImageJ.

    Lane 1: Buffer only. 

    Lane 2: Modified oligonucleotide (5 μM), 5’ Biotin-mN.mN.mN.mN.mN.[m6,6A]*.mN.mN.mN.mN.mN 3’

    Lane 3: Unmodified oligonucleotide (5 μM), 5’ Biotin-mN.mN.mN.mN.mN.[A]*.mN.mN.mN.mN.mN 3’

    N - equimolar mixture of (A/U/G/C)
    m - 2’O methyl protection
    * - phosphorothioate protection

     

    Blocking buffer and concentration: 5% NFDM/TBST

    Dilution buffer and concentration: TBST/0.1% Triton X-100/1 mM EDTA

  • ELISA - Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] (ab208198)
    ELISA - Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] (ab208198)

    Biotinylated m6,6A (modified), m6A (modified) and A (unmodified) oligonucleotides with the below sequence were coated onto wells of a 96 well plate. 

    Modified oligonucleotide (5 μM), 5’ Biotin-mN.mN.mN.mN.mN.[m6,6A]*.mN.mN.mN.mN.mN 3’

    Modified oligonucleotide (5 μM), 5’ Biotin-mN.mN.mN.mN.mN.[m6A]*.mN.mN.mN.mN.mN 3’

    Unmodified oligonucleotide (5 μM), 5’ Biotin-mN.mN.mN.mN.mN.[A]*.mN.mN.mN.mN.mN 3’

    N - equimolar mixture of (A/U/G/C)
    m - 2’O methyl protection
    * - phosphorothioate protection

    ELISA was performed on 1.0 µg/ml of antigen using ab208198 at a concentration range of 0.005-4.000 µg/ml, followed by Goat Anti-Rabbit IgG, (H+L), alkaline phosphatase conjugated secondary antibody at 1/2500 dilution.

References

ab208198 has not yet been referenced specifically in any publications.

Publishing research using ab208198? Please let us know so that we can cite the reference in this datasheet.

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