Product: METTL3 Antibody
Catalog: DF12020
Description: Rabbit polyclonal antibody to METTL3
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Xenopus
Mol.Wt.: 65-70 kDa,95kDa; 64kD(Calculated).
Uniprot: Q86U44
RRID: AB_2844825

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Zebrafish(91%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Xenopus(82%)
Clonality:
Polyclonal
Specificity:
METTL3 Antibody detects endogenous levels of total METTL3.
RRID:
AB_2844825
Cite Format: Affinity Biosciences Cat# DF12020, RRID:AB_2844825.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

adoMet-binding subunit of the human mRNA (N6-adenosine)-methyltransferase; IME4; M6A; Methyltransferase like protein 3; Methyltransferase-like protein 3; METTL3; mRNA (2'-O-methyladenosine-N(6)-)-methyltransferase; mRNA m(6)A methyltransferase; MT-A70; MTA70; MTA70_HUMAN; N6 adenosine methyltransferase 70 kDa subunit; N6-adenosine-methyltransferase 70 kDa subunit;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q86U44 MTA70_HUMAN:

Widely expressed at low level. Expressed in spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes.

Sequence:
MSDTWSSIQAHKKQLDSLRERLQRRRKQDSGHLDLRNPEAALSPTFRSDSPVPTAPTSGGPKPSTASAVPELATDPELEKKLLHHLSDLALTLPTDAVSICLAISTPDAPATQDGVESLLQKFAAQELIEVKRGLLQDDAHPTLVTYADHSKLSAMMGAVAEKKGPGEVAGTVTGQKRRAEQDSTTVAAFASSLVSGLNSSASEPAKEPAKKSRKHAASDVDLEIESLLNQQSTKEQQSKKVSQEILELLNTTTAKEQSIVEKFRSRGRAQVQEFCDYGTKEECMKASDADRPCRKLHFRRIINKHTDESLGDCSFLNTCFHMDTCKYVHYEIDACMDSEAPGSKDHTPSQELALTQSVGGDSSADRLFPPQWICCDIRYLDVSILGKFAVVMADPPWDIHMELPYGTLTDDEMRRLNIPVLQDDGFLFLWVTGRAMELGRECLNLWGYERVDEIIWVKTNQLQRIIRTGRTGHWLNHGKEHCLVGVKGNPQGFNQGLDCDVIVAEVRSTSHKPDEIYGMIERLSPGTRKIELFGRPHNVQPNWITLGNQLDGIHLLDPDVVARFKQRYPDGIISKPKNL

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Zebrafish
91
Xenopus
82
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q86U44 As Substrate

Site PTM Type Enzyme
S2 Acetylation
S2 Phosphorylation
K27 Ubiquitination
S30 Phosphorylation
R36 Methylation
S43 Phosphorylation
S48 Phosphorylation
S50 Phosphorylation
T54 Phosphorylation
T57 Phosphorylation
S64 Phosphorylation
T65 Phosphorylation
K132 Sumoylation
K132 Ubiquitination
S196 Phosphorylation
S200 Phosphorylation
S203 Phosphorylation
K215 Ubiquitination
S219 Phosphorylation
K235 Ubiquitination
K241 Ubiquitination
S243 Phosphorylation
K256 Ubiquitination
K263 Ubiquitination
K305 Ubiquitination
S315 Phosphorylation
S344 Phosphorylation
T348 Phosphorylation
S350 Phosphorylation
T356 Phosphorylation
K459 Ubiquitination
K480 Ubiquitination
K488 Ubiquitination
K513 Ubiquitination
K566 Ubiquitination
K576 Ubiquitination

Research Backgrounds

Function:

The METTL3-METTL14 heterodimer forms a N6-methyltransferase complex that methylates adenosine residues at the N(6) position of some RNAs and regulates various processes such as the circadian clock, differentiation of embryonic and hematopoietic stem cells, cortical neurogenesis, response to DNA damage, differentiation of T-cells and primary miRNA processing. In the heterodimer formed with METTL14, METTL3 constitutes the catalytic core. N6-methyladenosine (m6A), which takes place at the 5'-[AG]GAC-3' consensus sites of some mRNAs, plays a role in mRNA stability, processing, translation efficiency and editing. M6A acts as a key regulator of mRNA stability: methylation is completed upon the release of mRNA into the nucleoplasm and promotes mRNA destabilization and degradation. In embryonic stem cells (ESCs), m6A methylation of mRNAs encoding key naive pluripotency-promoting transcripts results in transcript destabilization, promoting differentiation of ESCs (By similarity). M6A regulates the length of the circadian clock: acts as an early pace-setter in the circadian loop by putting mRNA production on a fast-track for facilitating nuclear processing, thereby providing an early point of control in setting the dynamics of the feedback loop (By similarity). M6A also regulates circadian regulation of hepatic lipid metabolism. M6A regulates spermatogonial differentiation and meiosis and is essential for male fertility and spermatogenesis (By similarity). Involved in the response to DNA damage: in response to ultraviolet irradiation, METTL3 rapidly catalyzes the formation of m6A on poly(A) transcripts at DNA damage sites, leading to the recruitment of POLK to DNA damage sites. M6A is also required for T-cell homeostasis and differentiation: m6A methylation of transcripts of SOCS family members (SOCS1, SOCS3 and CISH) in naive T-cells promotes mRNA destabilization and degradation, promoting T-cell differentiation (By similarity). Inhibits the type I interferon response by mediating m6A methylation of IFNB. M6A also takes place in other RNA molecules, such as primary miRNA (pri-miRNAs). Mediates m6A methylation of Xist RNA, thereby participating in random X inactivation: m6A methylation of Xist leads to target YTHDC1 reader on Xist and promote transcription repression activity of Xist. M6A also regulates cortical neurogenesis: m6A methylation of transcripts related to transcription factors, neural stem cells, the cell cycle and neuronal differentiation during brain development promotes their destabilization and decay, promoting differentiation of radial glial cells (By similarity). METTL3 mediates methylation of pri-miRNAs, marking them for recognition and processing by DGCR8. Acts as a positive regulator of mRNA translation independently of the methyltransferase activity: promotes translation by interacting with the translation initiation machinery in the cytoplasm. Its overexpression in a number of cancer cells suggests that it may participate to cancer cell proliferation by promoting mRNA translation.

PTMs:

Sumoylation inhibits the N6-adenosine-methyltransferase activity. Sumoylation does not affect subcellular location or interaction with METTL14. Desumoylated by SENP1.

Subcellular Location:

Nucleus. Nucleus speckle. Cytoplasm.
Note: Colocalizes with speckles in interphase nuclei, suggesting that it may be associated with nuclear pre-mRNA splicing components (PubMed:9409616). In response to ultraviolet irradiation, colocalizes to DNA damage sites however, it probably does not bind DNA but localizes in the vicinity of DNA damage sites (PubMed:28297716).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Widely expressed at low level. Expressed in spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes.

Subunit Structure:

Heterodimer; heterodimerizes with METTL14 to form an antiparallel heterodimer that constitutes an active methyltransferase. Component of the WMM complex, a N6-methyltransferase complex composed of a catalytic subcomplex, named MAC, and of an associated subcomplex, named MACOM. The MAC subcomplex is composed of METTL3 and METTL14. The MACOM subcomplex is composed of WTAP, ZC3H13, CBLL1/HAKAI, VIRMA, and, in some cases of RBM15 (RBM15 or RBM15B). Interacts with NCBP1/CBP80. Interacts with EIF4E. Interacts with EIF3B.

Family&Domains:

Gate loop 1 and gate loop 2 regions are adjacent to the S-adenosyl-L-homocysteine-binding site and display large conformational changes upon ligand-binding. They may play an important role in adenosine recognition. The interface loop contributes to the heterodimer interaction.

Belongs to the MT-A70-like family.

References

1). MiR-29a-3p Improves Acute Lung Injury by Reducing Alveolar Epithelial Cell PANoptosis. Aging and Disease, 2022 (PubMed: 35656115) [IF=7.0]

Application: WB    Species: Mouse    Sample: lung tissue

Figure 2. METTL3-mediated m6A modification stabilizes the expression of miR-29a-3p. (A) The level of total m6A in the lung tissue of ALI mouse model. (B) m6A RIP detects the m6A level of pri-miR-29a in the lung tissue of ALI mouse model. (C) RT-qPCR detection of METL3 expression in the lung tissue of ALI mouse model. (D) Western blot analysis for the expression of METTL3 in the lung tissue of ALI mouse model. (E) Western blot analysis for the expression of METTL3 in A549 cells after siRNA transfection. (F) RT-qPCR analysis for the level of miR-29a-3p in A549 cells after METTL3 siRNA transfection. *** p <0.001.

2). METTL3 contributes to slow transit constipation by regulating miR‐30b‐5p/PIK3R2/Akt/mTOR signaling cascade through DGCR8. JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, 2022 (PubMed: 36068012) [IF=3.7]

3). METTL3/14 and IL‐17 signaling contribute to CEBPA‐DT enhanced oral cancer cisplatin resistance. ORAL DISEASES, 2023 (PubMed: 34807506) [IF=2.9]

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