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AB228860

Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] - BSA and Azide free

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Rabbit Recombinant Monoclonal N6, N6-dimethyladenosine (m6,6A) antibody. Carrier free. Suitable for IP, ELISA and reacts with Modified Nucleic Acid samples.

View Alternative Names

Di-m6A, N6, N6 dimethyladenosine

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ELISA - Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] - BSA and Azide free (AB228860)
  • ELISA

Lab

ELISA - Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] - BSA and Azide free (AB228860)

Biotinylated m6,6A (modified), m6A (modified) and A (unmodified) oligonucleotides with the below sequence were coated onto wells of a 96 well plate.

Modified oligonucleotide (5 μM), 5’ Biotin-mN.mN.mN.mN.mN.[m6,6A]*.mN.mN.mN.mN.mN 3’

Modified oligonucleotide (5 μM), 5’ Biotin-mN.mN.mN.mN.mN.[m6A]*.mN.mN.mN.mN.mN 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

ELISA was performed on 1.0 µg/ml of antigen using ab208198 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208198).

Immunoprecipitation - Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] - BSA and Azide free (AB228860)
  • IP

Supplier Data

Immunoprecipitation - Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] - BSA and Azide free (AB228860)

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

ab208198 (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.[m6,6A]*.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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208198).

All lanes:

Immunoprecipitation - Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44] (<a href='/en-us/products/primary-antibodies/n6-n6-dimethyladenosine-m66a-antibody-epr-19831-44-ab208198'>ab208198</a>)

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  • Unconjugated

    Anti-N6, N6-dimethyladenosine (m6,6A) antibody [EPR- 19831-44]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR- 19831-44

Isotype

IgG

Carrier free

Yes

Applications

IP, ELISA

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

Has been developed to discriminate between the modified base N6, N6-dimethyladenosine (m6,6A) and the unmodified counterpart Adenosine (A).

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "ELISA" : {"fullname" : "ELISA", "shortname":"ELISA"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Modified Nucleic Acid": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "ELISA-species-checked": "testedAndGuaranteed", "ELISA-species-dilution-info": "0.005-4 µg/mL", "ELISA-species-notes": "<p></p>" } } }
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Product details

ab228860 is the carrier-free version of ab208198.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

N6 N6-dimethyladenosine (m66A) also known as MMA is a modified nucleotide found in various RNA molecules. It possesses two methyl groups at the N6 position of the adenine base contributing to the stability and processing of RNA. The molecular mass of m66A is approximately 267.27 g/mol. This modification appears in a wide range of organisms from bacteria to humans indicating its importance across species. In human cells m66A is prevalent within the cytoplasm and nucleus where it plays a role in modulating RNA behavior.
Biological function summary

M66A can affect RNA's structural configuration and its interaction with other molecules. It is a part of the broader modification network within RNAs often found with other RNA modifications such as m6A and m1A. The presence of m66A influences RNA stability splicing and export which can alter the outcome of gene expression. In the context of gene expression regulation m66A interacts with multiple proteins that recognize and modify RNA structures. Consequently it forms part of the dynamic and responsive mechanisms that control cellular activities.

Pathways

M66A participates in the RNA metabolism and gene expression pathways. It associates with critical biological functions through interactions with proteins like METTL3 and FTO which regulate RNA methylation status. These pathways are important for processes such as translation regulation and RNA decay. METTL3 a methyltransferase adds methyl groups to RNA while FTO a demethylase removes them creating a balance in the modification states essential for proper cellular function.

M66A shows connections to several conditions including cancer and metabolic disorders. Changes in m66A levels might influence cellular transformation and tumor progression linking it to oncogenic processes. Specifically altered interactions with METTL3 have been observed in certain cancers due to its role in modifying RNA and affecting gene expression programs. Additionally m66A's involvement in metabolic regulation suggests a potential link to metabolic disorders wherein disruptions in RNA modifications may lead to impaired metabolism and related diseases.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

See full target information N6, N6-dimethyladenosine (m6,6A)
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Product promise

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For full details, please see our Terms & Conditions

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