Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR-19836-208] to 1-methyladenosine (m1A)
- Suitable for: IP, Dot blot, ELISA
- Reacts with: Human
Product nameAnti-1-methyladenosine (m1A) antibody [EPR-19836-208]
See all 1-methyladenosine (m1A) primary antibodies
DescriptionRabbit monoclonal [EPR-19836-208] to 1-methyladenosine (m1A)
SpecificityHas been developed to discriminate between the modified base 1-methyladenosine (m1A) and the unmodified counterpart Adenosine (A).
Tested applicationsSuitable for: IP, Dot blot, ELISAmore details
Species reactivityReacts with: Human, Synthetic fragment
Predicted to work with: Mouse, Rat, a wide range of other species
Chemical/ Small Molecule corresponding to 1-methyladenosine (m1A).
- IP: 5' Biotin-mN-mN-mN-mN-mN-[m1A]-mN-mN-mN-mN-mN 3'. ELISA: BSA-conjugated m1A-modified nucleotide.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped 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 bufferpH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
PurityProtein A purified
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.
|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.|
RelevanceN1-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.
- 1 methyladenosine antibody
- 1-methyladenosine antibody
- m1A antibody
Primary antibody dilution: 1/500
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated
Secondary antibody dilution: 1/20,000
Blocking buffer and dilution buffer: AdvanBlockTM Chemi Blocking buffer
Input: HeLa total RNA 0.5 µg per Dot
Competitive nucleosides: m1A, m2A, m2.2G
Exposure time: 37 seconds
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
ab208196 has been referenced in 1 publication.
- Grozhik AV et al. Antibody cross-reactivity accounts for widespread appearance of m1A in 5'UTRs. Nat Commun 10:5126 (2019). Human . PubMed: 31719534