重组Anti-1-methylguanosine (m1G)抗体[EPR19833-150] - BSA and Azide free

• Dot

Supplier Data

Dot Blot - Anti-1-methylguanosine (m1G) antibody [EPR19833-150] - BSA and Azide free (AB228858)

This data was developed using ab208199, the same antibody clone in a different buffer formulation.

Dot blot analysis of 1-methylguanosine (m1G) using ab208199 at 1/500 dilution, followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20,000 dilution (Left) or Methylene Blue (Right).

Lane 1 : HeLa total RNA

Lane 2 : Yeast tRNA

Exposure time : 8 seconds

• IP

Supplier Data

Immunoprecipitation - Anti-1-methylguanosine (m1G) antibody [EPR19833-150] - BSA and Azide free (AB228858)

This data was developed using ab208199, the same antibody clone in a different buffer formulation.

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

ab208199 (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.[m1G]*.mN.mN.mN.mN.mN 3'

Lane 3 : Unmodified oligonucleotide (5 μM), 5' Biotin-mN.mN.mN.mN.mN.[G]*.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

All lanes:

Immunoprecipitation - Anti-1-methylguanosine (m1G) antibody [EPR19833-150] - BSA and Azide free (ab228858)

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• ELISA

Supplier Data

ELISA - Anti-1-methylguanosine (m1G) antibody [EPR19833-150] - BSA and Azide free (AB228858)

This data was developed using ab208199, the same antibody clone in a different buffer formulation.

BSA-conjugated m1G (modified) and G (unmodified) nucleosides were coated onto wells of a 96 well plate. ELISA was performed on 1.0 μg/ml of antigen using ab208199 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.

• FRET

Supplier Data

FRET - Anti-1-methylguanosine (m1G) antibody [EPR19833-150] - BSA and Azide free (AB228858)

This data was developed using ab208199, the same antibody clone in a different buffer formulation.

A FRET-based assay was performed using 25 nm of each biotinylated oligo using ab208199 at a concentration range of 0.001-4.000 µg/ml.

• Dot

Supplier Data

Dot Blot - Anti-1-methylguanosine (m1G) antibody [EPR19833-150] - BSA and Azide free (AB228858)

This data was developed using ab208199, the same antibody clone in a different buffer formulation.

Dot blot analysis of 1-methylguanosine (m1G) using ab208199 at 1/500 dilution, followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20,000 dilution (Upper) or a Streptavidin-Alexa^®488 conjugate (Lower).

Lane 1 : Biotin-m1G RNA oligo

Lane 2 : Biotin-G RNA oligo

Lane 3 : Biotin-m2G RNA oligo

Lane 4 : Biotin-m22G RNA oligo

Exposure time 5 seconds.