Recombinant
RabMAb

Recombinant Anti-1-methyladenosine (m1A) antibody [EPR-19836-208] (ab208196)

Overview

  • Product name
    Anti-1-methyladenosine (m1A) antibody [EPR-19836-208]
    See all 1-methyladenosine (m1A) primary antibodies
  • Description
    Rabbit monoclonal [EPR-19836-208] to 1-methyladenosine (m1A)
  • Host species
    Rabbit
  • Specificity
    Has been developed to discriminate between the modified base 1-methyladenosine (m1A) and the unmodified counterpart Adenosine (A).
  • Tested applications
    Suitable for: IP, Dot blot, ELISAmore details
  • Species reactivity
    Reacts with: Human, Synthetic fragment
    Predicted to work with: Mouse, Rat, a wide range of other species
  • Immunogen

    Chemical/ Small Molecule corresponding to 1-methyladenosine (m1A).

  • Positive control
    • IP: 5' Biotin-mN-mN-mN-mN-mN-[m1A]-mN-mN-mN-mN-mN 3'. ELISA: BSA-conjugated m1A-modified nucleotide.
  • General notes

    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 long term. 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-19836-208
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab208196 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.

Use 0.2 µg.

Dot blot Use a concentration of 2 µg/ml.
ELISA Use a concentration of 0.005 - 4 µg/ml.

Target

  • Alternative names
    • 1 methyladenosine antibody
    • 1-methyladenosine antibody
    • m1A antibody

Images

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

    ab208196 (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.[m1A].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

  • BSA-conjugated m1A (modified) and A (unmodified) nucleosides were coated onto wells of a 96 well plate. ELISA was performed on 1.0 µg/ml of antigen using ab208196 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.

  • Dot blot of total RNA using ab208196 at 2 ug/mL. The Amersham Hybond N+ membrane was pre-spotted with 500, 250, 125, 63 and 32 ng/dot of HeLa total RNA and 500ng of an unmodified RNA probe. The membrane was then blocked with 5% BSA in TBS with 0.1% Tween-20. Followed by blotting with anti-m1A ab208196, or ab208196 together with 100uM of free m1A nucleoside in the same blocking solution, to inhibit m1A binding. A goat anti-rabbit HRP was used as the secondary antibody at 1:5000 dilution. Methylene blue stain was used to verify RNA loading.

References

ab208196 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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