Mettl3

Manufactured by ABclonal

Sourced in China

METTLER3 is a lab equipment product that functions as an enzyme involved in the post-transcriptional modification of RNA. It catalyzes the methylation of adenosine residues in target RNAs, which is a crucial step in the regulation of gene expression.

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Lab products found in correlation

Flag tag (flag), by Merck Group (2 mentions) Gapdh, by Proteintech (2 mentions) N cadherin, by Proteintech (1 mentions) Mettl16, by Cell Signaling Technology (1 mentions) Mettl14, by Proteintech (1 mentions) Histone h3, by Cell Signaling Technology (1 mentions) β catenin, by Proteintech (1 mentions) E cadherin, by ABclonal (1 mentions) Ab52757, by Abcam (1 mentions) Ab134175, by Abcam (1 mentions) Pd150606 d5946, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Sw480, by American Type Culture Collection (1 mentions) Sw620, by American Type Culture Collection (1 mentions) Dld 1, by American Type Culture Collection (1 mentions) Cyclin d1, by Abcam (1 mentions) Gapdh, by Abcam (1 mentions) Cyclin d1, by Proteintech (1 mentions) Pakt1 ser 473, by Cell Signaling Technology (1 mentions)

6 protocols using mettl3

Protein Expression and Methylation Analysis

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Reverse transcription-quantitative PCR (RT-qPCR) and western blot methods were described previously 20 . The oligos and primers used in this study were listed in table S1. Moreover, the primary antibodies used in this study were as follows: GAPDH (cat. no. 2118, 1:1,000; Cell Signaling Technology), histone H3 (cat. no. 4499t, 1/1,000; Cell Signaling Technology), METTL16 (cat. no. A15894, 1/1,000; ABclonal), E-cadherin (cat. no. 20874-1-AP, 1/10,000; Proteintech), N-cadherin (cat. no. 66219-1-Ig, 1/5,000; Proteintech), ZO-1 (cat. no. 21773-1-AP, 1/5,000; Proteintech), DVL2 (cat. no. 12037-1-AP, 1/5,000; Proteintech), β-catenin (cat. no. A19657, 1/1,000; ABclonal), Phospho-β-catenin-S45 (cat. no. AP0580, 1/1,000; ABclonal), METTL3 (cat. no. A8370, 1/1,000; ABclonal), and METTL14 (cat. no. 26158-1-AP, 1/1,000; Proteintech).

Yu L., Lu J., Xie N., Fang L., Chen S., Wu Y., Wang X, & Li B. (2023). Suppression of Wnt/β-catenin Signaling in PDAC via METTL16-mediated N6-methyladenosine Modification of DVL2. Journal of Cancer, 14(16), 2964-2977.

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Colorectal Cancer Cell Line Culture

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Human CRC cell lines SW480, SW620, DLD1, RKO were obtained from the American Type Culture Collection (ATCC). All cells were routinely cultured in RPMI 1640 (Invitrogen; 11875–093) or DMEM (Invitrogen; 11965–092) supplemented with 10% fetal bovine serum. All cells were incubated at 37 °C with 5% CO2 and 95% humidity. The following antibodies were used for western blotting and IHC: HSF1 (12972S, Cell Signaling Technology, CST; ab52757, Abcam); β-catenin (8480S, CST); β-actin (4970 L, CST); FLAG (F1804–1, Sigma); cyclinD1 (ab134175, Abcam); Cleaved PARP1 (9541-s, CST); METTL3 (a8370, Abclonal); GLS1 (ap8809b, Abgent). Pyrvinium (P0027), LiCl (793620), cycloheximide (R750107), chloroquine (C6628), MG132 (474790) and PD150606 (D5946) were purchased from Sigma-Aldrich.

Song P., Feng L., Li J., Dai D., Zhu L., Wang C., Li J., Li L., Zhou Q., Shi R., Wang X, & Jin H. (2020). β-catenin represses miR455-3p to stimulate m6A modification of HSF1 mRNA and promote its translation in colorectal cancer. Molecular Cancer, 19, 129.

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Comprehensive Protein Expression Analysis

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WB analysis was performed as previously described.24 (link) The primary antibodies were employed: CBX2 (Proteintech, China), CDK4/6 (Proteintech, China), cyclin D1 (Proteintech, China), Flag (Sigma-Aldrich, USA), Ha (Abmart, China), Mettl3 (Abclonal, China), and GAPDH (Abcam, England).

Sun R., Tu X., Chan S., Wang X., Ji Y., Wang Z., Yu Z., Zuo X., Zhang Q., Chen J., Han Q., Wang M., Zhao H., Zhang H, & Chen W. (2024). CBX2 Deletion Suppresses Growth and Metastasis of Colorectal Cancer by Mettl3-p38/ERK MAPK Signalling Pathway. Journal of Cancer, 15(8), 2123-2136.

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Proteomic Analysis of Cellular Signaling

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RIPA buffer (with PMSF and phosphatase inhibitor) was used to extract whole-cell lysates. Briefly, equal amounts of protein samples were separated via SDS-PAGE and transferred to polyvinylidene fluoride membranes. The targeted proteins were detected using different antibodies, including GAPDH (1:1000, Proteintech, USA), IGF2BP3 (1:1000, Proteintech, USA), AKT1 (1:1000, Cell Signaling Technology, USA), P-AKT1(ser473) (1:1000, Cell Signaling Technology, USA), KLK5 (1:1000, #abs136657, Abisin, Shanghai), PAR2 (1:1000, Abclonal Technology Co., Ltd.) and METTL3 (1:1000, Abclonal Technology Co., Ltd.).

Zhang J., Yang K., Bu J., Yan J., Hu X., Liu K., Gao S., Tang S., Gao L, & Chen W. (2022). IGF2BP3 promotes progression of gallbladder carcinoma by stabilizing KLK5 mRNA in N6-methyladenosine-dependent binding. Frontiers in Oncology, 12, 1035871.

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Western Blot Analysis of Protein Markers

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Protein sample separated on SDS-PAGE, transferred onto 0.45 μm PVDF membranes (Millipore), blocked with 3% BSA, incubated overnight at 4 °C with primary antibodies against METTL3 (ABclonal, China), PAI-1 (Abcam), YTHDF1 (Proteintech), JUN (Cell Signaling Technology), GAPDH (Proteintech), Fibrin (Merck, Germany), and detected using chemiluminescence. Densitometry analysis was performed using ImageJ software (National Institutes of Health) by measuring the band intensity for each group, and the data were normalized to GAPDH as an internal control.

Bai Q., Lu Y., Chen Y., Zhang H., Zhang W., Wu H, & Wen A. (2021). Endothelial METTL3 (Methyltransferase-Like 3) Inhibits Fibrinolysis by Promoting PAI-1 (Plasminogen Activator Inhibitor-1) Expression Through Enhancing Jun Proto-Oncogene N6-Methyladenosine Modification. Arteriosclerosis, Thrombosis, and Vascular Biology, 41(12), 2877-2889.

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Molecular Mechanisms of mRNA Modification

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ATF4 (rabbit, 11815S), p-mTOR (rabbit, 5536S), mTOR (rabbit, 2983S), p-p70S6K (rabbit, 9205S), p70S6K (rabbit, 2708S), pS6 (rabbit, 4857S), LC3B (rabbit, 3868S), and sequestosome 1 (SQSTM1, rabbit, 8025) monoclonal antibodies were purchased from Cell Signaling Technology (Shanghai, China). DDIT4 (rabbit, 10638-1-AP) and YTHDF2 (rabbit, 24744-1-AP) polyclonal antibodies were purchased from Proteintech. FTO (rabbit, ab126605), WTAP (rabbit, ab195380), ALKBH5 (rabbit, ab69325), and m⁶A (mouse, ab208577) antibodies were purchased from Abcam. METTL3 (rabbit, a8370), and METTL14 (rabbit, a8530) polyclonal antibodies were purchased from ABclonal. Compound 968 was purchased from Sigma-Aldrich (SML1327). Actinomycin D was purchased from Selleck Chemicals (S8964). Chloroquine (CQ) was obtained from Sigma-Aldrich (C6628). Meclofenamate sodium was purchased from MCE (HY-B1320). CB839 was purchased from Selleck Chemicals (S7655).

Han S., Zhu L., Zhu Y., Meng Y., Li J., Song P., Yousafzai N.A., Feng L., Chen M., Wang Y., Jin H, & Wang X. (2021). Targeting ATF4-dependent pro-survival autophagy to synergize glutaminolysis inhibition. Theranostics, 11(17), 8464-8479.

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