UcallM® m¹A Immunoprecipitation Kit

N1-methyladenosine (m¹A) is an important post-transcriptional modification of RNA, referring to methylation at the N1 position of the adenine base in RNA. Accumulating studies reveal that m¹A not only affects local RNA structure, protein-RNA interactions and translation of 5'UTR, but also exists in tRNA, rRNA, mRNA and mitochondrial transcripts. It plays a critical role in maintaining the correct structure and biological functions of these ncRNAs. The UcallM® m¹A Immunoprecipitation Kit adopts an optimized antibody-magnetic bead directional coupling strategy to efficiently capture m¹A-modified fragments and effectively reduce non-specific binding. After enriching m¹A-modified fragments in samples via immunoprecipitation, the fragments can be used as templates for quantitative detection of m¹A modification levels by m¹A meRIP-qPCR. In addition, libraries can be constructed for transcriptome sequencing to draw the genome-wide m¹A modification landscape. Kit Information

Experimental Workflow

Key Advantages 1.High-specific enrichment, m¹A antibody validated by multiple published articles 2.Optimized elution system & workflow, markedly reducing non-specific binding 3.Optimized magnetic bead-antibody coupling efficiency to ensure data reproducibility

Application 1.Tumor pathogenesis: e.g. dissecting pro-tumor pathways regulated by m¹A in cancers 2.Metabolic and immune regulation: study m¹A in T cell immune response, mitochondrial function and metabolic reprogramming 3.Stem cell fate regulation: investigate how dynamic m¹A modification modulates translation efficiency in stem cells Customer Publications PNAS: RNA modification regulates tumor cell glycolysis via ATP5D (DOI:10.1073/pnas.2119038119) This study verified via sequencing and functional assays that ATP5D is one of the core subunits of ATP synthase, participating in tumor cell glycolysis regulated by m¹A demethylase ALKBH3. The m¹A-modified exon of ATP5D increases binding to the YTHDF1/eRF1 complex to fine-tune its translational elongation, thereby promoting ribosome complex to release mRNA.

Figure 1-26. The study identified m¹A modification sites on ATP5D mRNA by m¹A-seq; m¹A-RIP-qPCR, SELECT-PCR and primer extension assays were used to verify m¹A modification at the A71 site of ATP5D mRNA.