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

  • Relevance

    N1-methyladenosine (m1A) is a RNA modification that has been reported in mRNA, tRNA, rRNA and lncRNA. It is found in bacteria, archaea and eukaryotes. The addition of the methyl group to the nitrogen at the 1st position of the adenosine base gives it a positive charge.
  • Alternative names

    • 1 methyladenosine antibody
    • 1-methyladenosine antibody
    • m1A antibody

Images

  • m1A was immunoprecipitated from 20 μg of HeLa (Human cervix adenocarcinoma epithelial cell) polyA+ RNA with 10 μg of ab208196 and 40 μL of Protein G dynabeads per sample (the IP buffer was 50mM Tris-HCl pH 7.4, 150mM NaCl, and 0.1% NP-40). The amount of m1A was quantified relative to the level of G by LC-MS/MS with electrospray ionization and in positive ionization mode, and compared to the level of m6A/G in the same samples. Error bars represent technical replicate injections of the same sample in mass spec.

  • 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.

  • 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

References

ab208196 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab208196.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up