Recombinant
RabMAb

Recombinant Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] - BSA and Azide free (ab228860)

Overview

  • Product name

    Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] - BSA and Azide free
    See all N6, N6-dimethyladenosine (m6,6A) primary antibodies
  • Description

    Rabbit monoclonal [EPR- 19831-44] to N6, N6-dimethyladenosine (m6,6A) - BSA and Azide free
  • 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

    Ab228860 is the carrier-free version of ab208198. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab228860 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    This product is a recombinant rabbit monoclonal antibody.

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

    Constituent: PBS
  • 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 ab228860 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.
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

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208198).

References

ab228860 has not yet been referenced specifically in any publications.

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