Diagenode

N6-methyladenosine (m6A) polyclonal antibody

Catalog Number
Format
Price
C15410208
50 μg
$380.00
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Polyclonal antibody raised in rabbit against N6-methyladenosine (m6A) conjugated to LPH.

LotA2125-0010
Concentration1.2 µg/µl
Species reactivityHuman, mouse, other (wide range): positive.
TypePolyclonal
PurityProtein G purified polyclonal antibody.
HostRabbit
Storage ConditionsStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
Storage BufferPBS containing 0.05% azide and 0.05% ProClin 300.
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
RIP/RIP-seq* 1-2 µg per IP Fig 1, 2, 3
Dot Blotting 1:400 Fig 4

*Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µg per IP.

  • Validation Data

    RIP

    Figure 1. RNA immunoprecipitation using the Diagenode antibody directed against m6A
    RNA Immunoprecipitation was performed on 40 µg HeLa total RNA spiked with 0.5 µg of an in vitro prepared transcript containing m6A nucleotides, using 2 µg of the Diagenode m6A antibody (Cat. No. C15410208). An equal amount of IgG was used as negative control. The immunoprecipitated RNA was subsequently analyzed by qRT-PCR with primers specific for the transcript and for an intergenic region, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    RNA immunoprecipitation

    Figure 2. RNA immunoprecipitation using the Diagenode antibody directed against m6A
    RNA Immunoprecipitation was performed on 40 µg HeLa total RNA fragmented to a mean size of ~500 bases. A titration consisting of 1, 2, 5 and 10 µg of antibody per RIP experiment was analyzed. IgG (2 µg/IP) was used as a negative IP control. Quantitative RT-PCR was performed with primers for 3’ UTR of the C1RL and SLC35E1 genes, and for exon 12 of the AFF1 gene, used as positive controls, and for exon 5 of the EIF2S3 gene, used as negative control.
    Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    A. RNA immunoprecipitation

    B. RNA immunoprecipitation

    C. RNA immunoprecipitation

    D. RNA immunoprecipitation

    Figure 3. RIP-seq results obtained with the Diagenode antibody directed against m6A
    RIP was performed with 2 µg of the Diagenode antibody against m6A (Cat. No. C15410208). The IP'd DNA was subsequently analysed on an Illumina HiSeq 4000. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 3 shows the signal distribution along the complete sequence of the human X-chromosome (figure 3A) and in three genomic regions surrounding the C1RL, SLC35E1 and AFF1 positive control genes (figure 3B, C and D).

    Dot blot

    Figure 2. Dot blot analysis using the Diagenode antibody directed against m6A
    To demonstrate the specificity of the Diagenode antibody against m6A (Cat. No. C15410208), a Dot Blot analysis was performed using synthetic oligonucleotides containing different modified bases. 150 and 50 pmol of the respective oligo’s were spotted on the membrane. The antibody was diluted 1:400 in PBS-T containing 10 % skimmed milk and 1% BSA. Figure 1 shows a high specificity of the antibody for the oligonucleotide with the N6-methyladenosine modification.

  •  実験手法
    DB
    Dot blotting Read more
    RIP
    RIP Read more
  •  資料
    Datasheet m6A C15410208 DATASHEET
    Datasheet description
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    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
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    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
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  •  Safety sheets
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  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: N6-methyladenosine (m6A) polyclonal antibody (Diagenode Cat# C15410208 Lot# A2125-0010). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Genomewide analysis of 6-methyladenine DNA in peripheral blood mononuclear cells of systemic lupus erythematosus.
    Zheng F, Tang D, Xu H, Xu Y, Dai W, Zhang X, Hong X, Liu D, Dai Y
    AIM: The aim of this paper is to explore the expression of 6-methyladenine (6mA) DNA and to elucidate its gene regulation role in systemic lupus erythematosus (SLE). METHODS: Twenty SLE patients and 20 normal control healthy individuals (HCs) were included in this study. Genomic DNA was isolated from peripheral bloo...

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