> Chemicals & Drugs > Amino Acid > METTL3 protein, human

METTL3 protein, human

METTL3 (Methyltransferase Like 3) is a protein that plays a crucial role in the methylation of RNA.
It is a member of the N6-adenosine-methyltransferase (MTase) complex and is responsible for the addition of methyl groups to adenosine residues in various RNA species, including mRNA, tRNA, and snRNA.
This post-transcriptional modification can influence RNA stability, localization, and translation, making METTL3 an important regulator of gene expression.
Researchers can explore the latest METTL3 protein research using PubCompare.ai's AI-driven optimization platform, which helps locate protocols from literature, preprints, and patents, and uses AI comparisons to identify the best protocols and products for METTL3 experiments.
This can enhance reproducibility and streamline research effoorts, helping scientists gain deeper insights into the biological functions of this key methyltransferase.

Most cited protocols related to «METTL3 protein, human»

M6A Modification Patterns Analysis

Cited 26 times

Owing to the few m⁶A regulators detected by Illumina HumanRef-8 WG-DASL v3.0 platform, we did not include GSE26253 cohort for clustering analysis. A total of 21 regulators were extracted from five integrated GEO datasets for identifying different m⁶A modification patterns mediated by m⁶A regulators. These 21 m⁶A regulators included 8 writers (METTL3, METTL14, RBM15, RBM15B, WTAP, KIAA1429, CBLL1, ZC3H13), 2 erasers (ALKBH5, FTO) and 11 readers (YTHDC1, YTHDC2, YTHDF1, YTHDF2, YTHDF3, IGF2BP1, HNRNPA2B1, HNRNPC, FMR1, LRPPRC, ELAVL1). Unsupervised clustering analysis was applied to identify distinct m⁶A modification patterns based on the expression of 21 m⁶A regulators and classify patients for further analysis. The number of clusters and their stability were determined by the consensus clustering algorithm [25 (link)]. We used the ConsensuClusterPlus package to perform the above steps and 1000 times repetitions were conducted for guaranteeing the stability of classification [26 (link)].

Zhang B., Wu Q., Li B., Wang D., Wang L, & Zhou Y.L. (2020). m6A regulator-mediated methylation modification patterns and tumor microenvironment infiltration characterization in gastric cancer. Molecular Cancer, 19, 53.

Publication 2020

METTL3 protein, human METTL14 protein, human Patients RBM15 protein, human

In vitro Methyltransferase Activity Assay

Cited 17 times

In vitro methyltransferase activity assay was performed in a standard 50 μL of reaction mixture containing the following components: 0.15 nmol RNA probe, 0.15 nmol each recombinant protein (single METTL3, METTL14, WTAP, or their combinations with a molar ratio of 0.15 nmol/0.15 nmol for two components, 0.8 mM d₃-SAM, 80 mM KCl, 1.5 mM MgCl₂, 0.2 U μL⁻¹ RNasin, 10 mM DTT, 4% glycerol, and 15 mM HEPES (pH 7.9). Prior to the reaction, the RNA probes were annealed with a program of (i) 90 °C for 3 min, and (ii) −2 °C/cycle for 40 cycles within 30 min.
The reaction was incubated at 16 °C for 12 h. The resultant RNA was recovered by phenol/chloroform (low pH) extraction followed by ethanol precipitation, and digested by nuclease P1 and alkaline phosphatase for QQQ LC-MS/MS analysis. The nucleosides were quantified by using the nucleoside to base ion mass transitions of 285 to 153 (d₃-m⁶A) and 284 to 152 (G). G served as an internal control to calculate the amount of RNA probe in each reaction mixture.

Liu J., Yue Y., Han D., Wang X., Fu Y., Zhang L., Jia G., Yu M., Lu Z., Deng X., Dai Q., Chen W, & He C. (2013). A METTL3-METTL14 complex mediates mammalian nuclear RNA N6-adenosine methylation. Nature chemical biology, 10(2), 93-95.

Publication 2013

Alkaline Phosphatase Biological Assay Chloroform Ethanol Glycerin HEPES Magnesium Chloride Methyltransferase METTL3 protein, human METTL14 protein, human Molar Nucleosides Phenol Recombinant Proteins RNA Probes Tandem Mass Spectrometry

METTL3/14/4 and WTAP Knockdown Assay

Cited 9 times

Human HeLa cell line was grown in DMEM (Gibco, 11965) media supplemented with 10% FBS and 1% 100× Pen Strep (Gibco, 15140). Human 293FT cell line was grown in DMEM (Gibco, 11995) media supplemented with 10% FBS and 1% 100× Pen Strep. METTL3, METTL14, METTL4, and WTAP siRNAs were purchased from QIAGEN with sequences shown in Supplementary Table 1. Transfection was achieved by using Lipofectamine RNAiMAX (Invitrogen) for siRNA, or Lipofectamine 2000 (Invitrogen) for the plasmid following the manufacturer’s protocols.

Publication 2013

Cell Lines HeLa Cells Homo sapiens Lipofectamine lipofectamine 2000 METTL3 protein, human METTL14 protein, human Plasmids RNA, Small Interfering Streptococcal Infections Transfection

Transcriptome Profiling and m6A-Related lncRNAs in KIRC

Cited 7 times

As the flow chart of the study shown in Figure S1, we downloaded Transcriptome profiling data in fragment per kilobase method (FPKM) format of 530 KIRC patients from TCGA data portal (https://portal.gdc.cancer.gov/). Subsequently, these data were collated and annotated, and then collapsed into protein-coding genes and long non-coding RNAs employing the Ensembl human genome browser (http://asia.ensembl.org/info/data/index.html) using the Perl program (16 (link)). And 14,142 lncRNAs were identified. Then, the differential analysis of these lncRNAs was performed by the “limma” package in R 4.0.3 (logFC > 1 or<-1, p < 0.05), and 4,492 significantly differential lncRNAs were identified. In addition, 35 m6A-related genes were obtained from published articles (8 (link), 17 (link)), and the expression matrixes were extracted from transcriptome profiling datasets, including regulators on writers [KIAA1429 (VIRMA), METTL3, METTL14, WTAP, RBM15, RBM15B, METTL16, ZC3H13, and PCIF1], readers [TRMT112, ZCCHC4, NUDT21 (CPSF5), CPSF6, CBLL1 (HAKAI), SETD2, HNRNPC, HNRNPG (RBMX), HNRNPA2B1, IGF2BP1, IGF2BP2, IGF2BP3, YTHDC1, YTHDF1, YTHDF2, YTHDF3, YTHDC2, SRSF3, SRSF10, XRN1, FMR1 (FMRP), NXF1, and PRRC2A], and erasers (FTO, ALKBH5, and ALKBH3). The differential analysis was also performed by the “limma” package in R software and 25 m6A-related genes were confirmed to be significantly different (p < 0.05, Figure S2). Then, Pearson correlation analysis between these lncRNAs and 25 m6A-related genes was performed, and 753 m6A-related lncRNAs were identified (cor > 0.5 or <−0.5, p < 0.05). The clinicopathological data were downloaded from the TCGA dataset, excluding those with survival time <30 days or unknown (n = 17), and those with unclear specific information including stage (n = 3), tumor grade (n = 3), and AJCC M stage (n = 3). Subsequently, we merged lncRNAs expression data with clinical data. Ultimately, a total of 505 cases were included in the study.

Yu J., Mao W., Sun S., Hu Q., Wang C., Xu Z., Liu R., Chen S., Xu B, & Chen M. (2021). Identification of an m6A-Related lncRNA Signature for Predicting the Prognosis in Patients With Kidney Renal Clear Cell Carcinoma. Frontiers in Oncology, 11, 663263.

Publication 2021

Division Phase, Cell Fragile X Mental Retardation Protein Gene Products, Protein Genes Genome, Human hSet2 protein, human IGF2BP2 protein, human IGF2BP3 protein, human Malignant Neoplasms METTL3 protein, human METTL14 protein, human Neoplasms RBM15 protein, human RNA, Long Untranslated SRSF3 protein, human XRN1 protein, human

Catalytic Dead Mettl3 in Naive T Cells

Cited 7 times

The WT and catalytic dead Mettl3 genes were cloned into N103 plasmid in Chang lab. To create a catalytic dead mutant Mettl3, we inserted the mutations D395A and W398A (DPPW catalytic motif of METTL3) based on published data, which show that those two residues are absolutely required for METTL3 activity ^{35 (link)–37 (link)}. Mettl3 KO naïve T cells were isolated from Mettl3 KO mice with StemCell Naïve T cell purification kit, then electroporated by nucleofection by exactly following the Amaxa mouse T cell nucleofector kit manual (Lonza). 4 hours after nucleofection in 37°C incubator, the cells were transferred into Rag2^−/− receipt mice by intravenous injection (1 million cells per mice). 4 weeks after transfer, the cells from spleen, periphery lymph nodes, and mesenteric lymph nodes were analyzed by FACS for cell numbers, and naïve Cell marker CD45RB. The CD4+ T cells were isolated from lymph nodes using StemCell CD4+ T cell kit, and total RNA was isolated from the CD4+ T cells using Direct-zol RNA microprep columns (Zymo Research). RT-qPCR was performed using the isolated RNAs. Each group has at least 5 mice to ensure statistical significance.

Li H.B., Tong J., Zhu S., Batista P.J., Duffy E.E., Zhao J., Bailis W., Cao G., Kroehling L., Chen Y., Wang G., Broughton J.P., Chen Y.G., Kluger Y., Simon M.D., Chang H.Y., Yin Z, & Flavell R.A. (2017). m6A mRNA methylation controls T cell homeostasis by targeting IL-7/STAT5/SOCS pathway. Nature, 548(7667), 338-342.

Publication 2017

Catalysis CD4 Positive T Lymphocytes Cells Genes Mesentery METTL3 protein, human Mus Mutation Nodes, Lymph Plasmids RAG2 protein, human RNA Spleen Stem Cells T-Lymphocyte

Most recents protocols related to «METTL3 protein, human»

Analyzing Circular RNA Methylation in ARC

The circRNA-seq data and MeRIP-seq data were downloaded from the GEO database under accession number GSE153722 (six ARC vs. six normal). The differential expression analysis of circRNA-seq and MeRIP-seq was performed using the R package DESeq under the cut-off criteria: —log2FC—≥ 1. The m6A-enriched circRNA in the normal control and ARC samples were analyzed. The intersection of differentially expressed circRNAs and differentially methylated m6A-enriched circRNAs served as candidate circRNAs.
Additionally, the RNA-protein binding site between has_circ_0007905 and METTL3 was predicted using the RBPsuite software (http://www.csbio.sjtu.edu.cn/bioinf/RBPsuite/). The potential m6A modification sites of has_circ_0007905 were predicted using SRAMP software (http://www.cuilab.cn/sramp).

Li R., Zhu H., Li Q., Tang J., Jin Y, & Cui H. (2023). METTL3-mediated m6A modification of has_circ_0007905 promotes age-related cataract progression through miR-6749-3p/EIF4EBP1. PeerJ, 11, e14863.

Publication 2023

METTL3 protein, human RNA, Circular RNA-Binding Proteins

Western Blot Analysis of Protein Levels

HLE-B3 cells were harvested and lysed using a lysis buffer, and the protein concentration of cell extracts was quantified with a BSA kit. Next, appropriately 20 µg protein was loaded on 10 SDS-PAGE gels and transferred into PVDF membranes, followed by blocking with TBST solution containing 5% skim milk at 4 °C for 3 h. Membranes were incubated with primary antibodies at 4 °C overnight and then incubated with secondary antibodies of Goat Anti-Mouse IgG H&L (HRP) (1:1000, ab205719; Abcam, Cambridge, UK) and Goat Anti-Rabbit IgG H&L (HRP) (1:20000, ab6721; Abcam, Cambridge, UK). Finally, the protein bands were imaged using the Bio-Rad ChemiDoc XRS system. Primary antibodies, including GAPDH (1:2000, 60004-1-Lg; Proteintech, Rosemont, IL, USA), METTL3 (1:1000, 15073-I-AP; Proteintech), and EIF4EBP1 (1:2000, ab32024; Abcam, Cambridge, UK), were used.

Publication 2023

anti-IgG Antibodies Buffers Cardiac Arrest Cell Extracts GAPDH protein, human Gels Goat METTL3 protein, human Milk, Cow's Mus polyvinylidene fluoride Proteins Rabbits SDS-PAGE Tissue, Membrane

Western Blotting of Epigenetic Regulators

Cells were lysed in RIPA buffer (1% Triton X-100, 20 mM Na2PO4, 150 mM NaCl (pH 7.4)) containing PMSF and Phosphatase Inhibitor Cocktail (Roche). Subsequently, a BCA assay (Thermo Fisher Scientific) was used to determine the protein concentrations. Proteins were separated by SDS-PAGE gels and transferred onto nitrocellulose membranes (Millipore). The membranes were placed in TBST (10 mM Tris-HCl (pH 7.4), 150 mM NaCl, and 0.1% Tween-20) containing 5% nonfat milk for 1 h at room temperature. Primary antibodies were diluted in TBST containing 5% BSA and used at the indicated concentrations: rabbit anti-acetyl-α-tubulin (1:1,000, 5335, Cell Signaling Technology), rabbit anti-EZH2 (1:1,000, 5246, Cell Signaling Technology), rabbit anti-H3K27me3 (1:1,000, 9733, Cell Signaling Technology), rabbit anti-DNMT3A (1:1,000, ab2850, Abcam), rabbit anti-YTHDF1 (1:1,000, 17479-1-AP, 18810, Proteintech), rabbit anti-METTL3 (1:1,000, ab18810, Abcam), and mouse anti-β-actin (1:5,000, Sigma). The membranes were incubated with primary antibodies overnight at 4°C, washed with TBST 4 times and incubated with HRP-conjugated anti-mouse IgG (1:10,000, 7076, Cell Signaling Technology) or anti-rabbit IgG (1:10,000, 7074, Cell Signaling Technology) diluted in TBST at room temperature for 1 h. After 4 final washes with TBST, the membranes were developed by using ECL and visualized using Tanon 5500 or Amersham Imager 680.

Yin H., Ju Z., Zheng M., Zhang X., Zuo W., Wang Y., Ding X., Zhang X., Peng Y., Li J., Yang A, & Zhang R. (2023). Loss of the m6A methyltransferase METTL3 in monocyte-derived macrophages ameliorates Alzheimer’s disease pathology in mice. PLOS Biology, 21(3), e3002017.

Publication 2023

Actins alpha-Tubulin anti-IgG Antibodies Biological Assay Buffers Cells EZH2 protein, human Gels IGG-horseradish peroxidase METTL3 protein, human Milk, Cow's Mus Nitrocellulose Phosphoric Monoester Hydrolases Proteins Rabbits Radioimmunoprecipitation Assay SDS-PAGE Sodium Chloride Tissue, Membrane Triton X-100 Tromethamine Tween 20

Mettl3 and Fto Overexpression in Neurons

The Mettl3 and Fto cDNA plasmid was constructed by Genechem Co., LTD (Shanghai, China). 2 µg lentivirus was used to transfect neuron with transfection reagents, according to the manufacturers’ instruction for 12 h. At 12 h post-infection, the medium was replaced with the neuron maintenance medium. The gene expression level was analysed by Western blot.

Zhang C., Jian H., Shang S., Lu L., Lou Y., Kang Y., Bai H., Fu Z., Lv Y., Kong X., Li X., Feng S, & Zhou H. (2023). Crosstalk between m6A mRNAs and m6A circRNAs and the time-specific biogenesis of m6A circRNAs after OGD/R in primary neurons. Epigenetics, 18(1), 2181575.

Publication 2023

DNA, Complementary Gene Expression Infection Lentivirus METTL3 protein, human Neurons Plasmids Transfection Western Blot

Quantification of RNA Methylation Regulators

After the neurons were washed with PBS, RIPA lysis buffer (Merck Millipore, Cat. No. 20–188) containing protease inhibitors (Roche, Cat. No. 5,892791,001) was added to fully lyse the cells on ice for 30 minutes, followed by centrifugation at 12,000 X g at 4°C for 10 minutes. The supernatant was collected, and the PierceTM BCA Protein Assay Kit (Thermo Scientific, Cat. No. 23225) was used to determine the protein concentration. Finally, the protein was denatured at 100°C for 10 minutes. 20ug of protein were analysed by immunoblotting using an anti-Mettl3 antibody (Abcam Cat. No. ab195352), an anti-Fto antibody (Abcam Cat. No. ab280081), with an anti-β-actin antibody (Abcam Cat. No. ab8226) used as an internal control.

Publication 2023

Actins Antibodies, Anti-Idiotypic Biological Assay Buffers Centrifugation METTL3 protein, human Neurons Protease Inhibitors Proteins Radioimmunoprecipitation Assay

Top products related to «METTL3 protein, human»

Ab195352 by Abcam

Sourced in United Kingdom, United States, China

Ab195352 is a lab equipment product from Abcam. It is a device used for the detection and measurement of specific molecules or analytes in a sample.

Lipofectamine 3000 by Thermo Fisher Scientific

Sourced in United States, China, Germany, Japan, United Kingdom, France, Canada, Italy, Australia, Switzerland, Denmark, Spain, Singapore, Belgium, Lithuania, Israel, Sweden, Austria, Moldova, Republic of, Greece, Azerbaijan, Finland

Lipofectamine 3000 is a transfection reagent used for the efficient delivery of nucleic acids, such as plasmid DNA, siRNA, and mRNA, into a variety of mammalian cell types. It facilitates the entry of these molecules into the cells, enabling their expression or silencing.

Lipofectamine 2000 by Thermo Fisher Scientific

Sourced in United States, China, Germany, United Kingdom, Canada, Japan, France, Italy, Switzerland, Australia, Spain, Belgium, Denmark, Singapore, India, Netherlands, Sweden, New Zealand, Portugal, Poland, Israel, Lithuania, Hong Kong, Argentina, Ireland, Austria, Czechia, Cameroon, Taiwan, Province of China, Morocco

Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.

Pvdf membrane by Merck Group

Sourced in United States, Germany, China, United Kingdom, Morocco, Ireland, France, Italy, Japan, Canada, Spain, Switzerland, New Zealand, India, Hong Kong, Sao Tome and Principe, Sweden, Netherlands, Australia, Belgium, Austria

PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.

Trizol reagent by Thermo Fisher Scientific

Sourced in United States, China, Japan, Germany, United Kingdom, Canada, France, Italy, Australia, Spain, Switzerland, Netherlands, Belgium, Lithuania, Denmark, Singapore, New Zealand, India, Brazil, Argentina, Sweden, Norway, Austria, Poland, Finland, Israel, Hong Kong, Cameroon, Sao Tome and Principe, Macao, Taiwan, Province of China, Thailand

TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.

Anti mettl3 by Abcam

Sourced in United States, United Kingdom

Anti-METTL3 is a primary antibody that detects the expression of METTL3, a methyltransferase enzyme involved in N6-methyladenosine (m6A) modification of RNA. This antibody can be used in various research applications to study the role of METTL3 and m6A modification in cellular processes.

Mettl3 by Abcam

Sourced in United States, United Kingdom

METTLER3 is a protein that catalyzes the addition of methyl groups to RNA molecules. It plays a role in the post-transcriptional modification of RNA.

Mettl3 by Proteintech

Sourced in United States, China

METTLER3 is a protein that catalyzes the addition of methyl groups to RNA molecules. It plays a role in mRNA processing and stability.

Bca protein assay kit by Beyotime

Sourced in China, United States, Germany, Puerto Rico, United Kingdom, Switzerland, Japan, Sweden

The BCA protein assay kit is a colorimetric-based method for the quantitative determination of total protein concentration in a sample. It uses bicinchoninic acid (BCA) to detect and quantify the presence of protein.

Dual luciferase reporter assay system by Promega

Sourced in United States, China, Germany, United Kingdom, Switzerland, Japan, France, Italy, Spain, Austria, Australia, Hong Kong, Finland

The Dual-Luciferase Reporter Assay System is a laboratory tool designed to measure and compare the activity of two different luciferase reporter genes simultaneously. The system provides a quantitative method for analyzing gene expression and regulation in transfected or transduced cells.

More about "METTL3 protein, human"

Explore the latest research on the METTL3 (Methyltransferase Like 3) protein, a key regulator of gene expression through its role in RNA methylation.
METTL3 is a member of the N6-adenosine-methyltransferase (MTase) complex, responsible for adding methyl groups to adenosine residues in various RNA species, including mRNA, tRNA, and snRNA.
This post-transcriptional modification can influence RNA stability, localization, and translation, making METTL3 an important player in cellular processes.
Leverage PubCompare.ai's AI-driven optimization platform to streamline your METTL3 research efforts.
Locate relevant protocols from literature, preprints, and patents, and use AI-powered comparisons to identify the best protocols and products for your experiments.
This can enhance reproducibility and help you gain deeper insights into the biological functions of this crucial methyltransferase.
Key tools and reagents that can support your METTL3 research include Ab195352 antibody, Lipofectamine 3000 and Lipofectamine 2000 for transfection, PVDF membranes for Western blotting, TRIzol reagent for RNA extraction, and the Dual-Luciferase Reporter Assay System for studying METTL3-mediated transcriptional regulation.
Utilize the BCA protein assay kit to quantify protein levels in your samples.
Explore the latest advancements in METTL3 research and optimize your experimental workflow with the help of PubCompare.ai's innovative tools and resources.
Enhance reproducibility, save time, and uncover new insights about this key methyltransferase and its role in gene expression.