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Anti-N6-methyladenosine (m6A) [17-3-4-1]
RECENTLY UPDATED

Invented by Dr Rupert Fray at University of Nottingham

Info

Catalogue Number 152130
Applications
RECENTLY UPDATED
IP DB
Antigen/Gene or Protein Targets N6-methyladenosine-5'-mono-phosphate
Reactivity Human, Mouse and Saccharomyces cerevisiae
Relevance
RECENTLY UPDATED
Monoclonal antibody which binds to m6A modification found in RNA and allows for analysis of methylated transcriptomes. This can be used to investigate how m6A modifications regulates gene expression.

Background and Research Application
N6-Methyladenosine (m6A) is an abundant modification found in mRNA, tRNA, snRNA, as well as long non-coding RNA, in all species. RNA adenosine methylation is catalysed by a multicomponent complex composed of METTL3/MT-A70, METTL14, and WTAP in mammals. METTL3 & METTL14 are responsible for the methyltransferase activity of the complex, and WTAP mediates substrate recruitment.
This antibody was created to identify m6A containing messages within RNA and helped identify the transcripts of three key, early regulators of meiosis, IME1, IME2 and IME4 itself as being methylated in RNA transcripts. Anti-N6-methyladenosine helped narrow down the position of m6A in IME2 as towards the 3’ end.
Anti-N6-methyladenosine will allow for analysis of methylated transcriptomes, and could aid understanding in how the m6A base modification regulates gene expression.
Host Mouse
Immunogen Hapten N6-methyladenosine-5'-mono-phosphate conjugated to BSA of all N6-methyladenosine
Subclass IgG1 lambda
Myeloma Used Sp2/0-Ag14
Recommended Growing Conditions DMEM with fetal calf serum (15%), L-Glutamine (2 x 10-3 M), stable Glutamine (2 x 10-3 M), non-essential amino acid solution MEM, 2-mercaptoethanol (5 x 10-5 M)
Strain Balb/c
Notes
RECENTLY UPDATED
Production Details
Purified using multi-step affinity chromatography with protein A.

Storage Conditions
Store at -20 degrees frozen. Avoid repeated freeze/thaw cycles.

Points of Interest
It was shown that mutations in the m6A demethylase gene FTO, which cause a decrease of m6A levels, are associated with an increased risk for obesity and type 2 diabetes.

This antibody recognises N6-methyladenosine in both modified RNA AND DNA (see associated data).

Application Note: For immunoprecipitation the best results are obtained by pre-binding the antibody to the beads before washing off unbound immunoglobulin. It is recommended to start with an excess of antibody in order to ensure the Protein G beads are saturated.

IP protocol was derived from Dominissini et al., 2013 Nat Protoc. 8(1):176-89.

Concentration
1mg/ml as standard
Research Area Epigenetics & Nuclear Signalling, Metabolism

References

There are 7 reference entries for this reagent.

View All References

References: 7 entries
RECENTLY UPDATED

Lan et al. 2019. Mol Cancer. 18(1):186. PMID: 31856849.

Barbieri et al. 2017. Nature. 552(7683):126-131. PMID: 29186125.

IP

Promoter-bound METTL3 maintains myeloid leukaemia by m6A-dependent translation control.

Europe PMC ID: 29186125

Lannes et al. 2016. Sci Rep. 6:31563. PMID: 27539363.

N6-Methyladenosine in Nuclear RNA is a Major Substrate of the Obesity-Associated FTO. Jia et al. 2011. Nat Chem Biol. 7(12):885-7. PMID: 22002720.

DB

Bodi et al. 2010. Nucleic Acids Res. 38(16):5327-35. PMID: 20421205.

Yeast targets for mRNA methylation.

Europe PMC ID: 20421205


Add a reference

References: 7 entries
RECENTLY UPDATED

Lan et al. 2019. Mol Cancer. 18(1):186. PMID: 31856849.

Barbieri et al. 2017. Nature. 552(7683):126-131. PMID: 29186125.

IP

Promoter-bound METTL3 maintains myeloid leukaemia by m6A-dependent translation control.

Lannes et al. 2016. Sci Rep. 6:31563. PMID: 27539363.

N6-Methyladenosine in Nuclear RNA is a Major Substrate of the Obesity-Associated FTO. Jia et al. 2011. Nat Chem Biol. 7(12):885-7. PMID: 22002720.

DB

Bodi et al. 2010. Nucleic Acids Res. 38(16):5327-35. PMID: 20421205.

Yeast targets for mRNA methylation.


Add a reference