ANTI-MCM5 (C005B-255608)

General description

N6-adenosine-methyltransferase catalytic subunit (UniProt: Q86U44; also known as EC:2.1.1.348, Methyltransferase-like protein 3, hMETTL3, N6-adenosine-methyltransferase 70 kDa subunit, MT-A70) is encoded by the METTL3 (also known as MTA70) gene (Gene ID: 56339) in human. METTL3 heterodimerizes with METTL14 to form an antiparallel heterodimer, which constitutes an active methyltransferase that methylates adenosine residues at the N6 position of some RNAs and regulates various cellular processes, including differentiation of embryonic and hematopoietic stem cells, cortical neurogenesis, differentiation of T-cells, and primary miRNA processing. In response to UV caused DNA damage, METTL3 rapidly catalyzes the formation of m6A on poly(A) transcripts at DNA damage sites that leads to the recruitment of POLK to these sites. It is localized both in the cytoplasm and in the nucleus. It colocalizes with speckles in interphase nuclei. In response to UV irradiation, it is shown to colocalize to DNA damage sites. Its methyltransferase activity is inhibited by sumoylation. However, sumoylation does not affect its subcellular location or interaction with METTL14. Overexpression of METTL3 is reported in several human cancers and its knockdown is known to promote apoptosis. METTL3 is reported to be strongly and directly phosphorylated by ERK2 at serine 43, 50, and 525 and to a lesser extent by p38 and JNK. METTL3 phosphorylation increases its USP5-mediated deubiquitination that leads to its stabilization. Phosphorylation of METTL3 is reported to promote murine embryonic stem cell differentiation. This polyclonal antibody specifically recognizes METTL3 phosphorylated at serine 43 and does not detect mutant forms where serine is replaced by alanine residues. (Ref.: Sun, H-L., et al. (2020). Mol. Cell. 80(4); 633-647; Wang, P., et al. (2016). Mol. Cell. 63(2); 306-317; Lin, S., et al. (2016). Mol. Cell. 62(3); 335-345; Wang, X. et al. (2014). Nature. 505(7481); 117-120).

Specificity

This rabbit polyclonal antibody specifically detects METTL3 phosphorylated on serine 43. It targets an epitope within 17 amino acids from the N-terminal region.

Immunogen

KLH-conjugated linear peptide corresponding to 17 amino acids surrounding phosphoserine 43 from the N-terminal region of human METTL3.

Application

Quality Control Testing

Evaluated by Western Blotting in lysates from HEK293T cells co-transfected with HER2 and Myc-tagged METTL3.

A 1:1,000 dilution of this antibody detected METTL3 phosphorylated on serine 43 in lysates from HEK293T cells co-transfected with HER2 and Myc-tagged METTL3, but not in lysates from cells transfected only with Myc-tagged METTL3.

Tested Applications

Western Blotting Analysis: A representative lot detected phospho-METTL3 (Ser43) in Western Blotting applications (Lung Sun, Hui., et. al. (2020). Mol Cell. 80(4): 633-647).

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected Phospho-METTL3 (Ser 43) in HEK293T cells co-transfected HER2 and Myc-tagged METTL3 (Data courtesy of Dr Chuan He & Dr Hui-Lung Sun at the University of Chicago).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Anti-Phospho-METTL3 (Ser 43), Cat. No. ABE2611, is a rabbit polyclonal antibody that detects METTL3 phosphorylated on serine 43 and is tested for use in Western Blotting.

Physical form

Purified rabbit polyclonal antibody in PBS with 0.25% BSA and 0.02% sodium azide

Storage and Stability

Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.