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.
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 bufferPreservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 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.|
- 1 methyladenosine antibody
- 1-methyladenosine antibody
- m1A antibody
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.
ab208196 has not yet been referenced specifically in any publications.