Human cells were cultured in standard cell culture Dulbecco's modified Eagle's medium at 37 °C in a humidified incubator with 5% CO2(v/v). The human METTL3 gene was cloned into pCMV-Myc (Invitrogen), S-protein/FLAG/SBP (streptavidin-binding protein) triple-tagged destination vector48 (link) or pGEX-5×-2 (GE healthcare). The human WTAP gene was cloned into p3XFLAG-CMV-14 (Sigma) or pProEX-HTb (Invitrogen). The human METTL14 gene was cloned into pcDNA3-HA (Invitrogen). The following antibodies were used: rabbit-anti-METTL3 (Abcam), mouse-anti-METTL3 (Abnova), mouse-anti-WTAP (Santa Cruz), rabbit-anti-METTL14 (Atlas), rabbit-anti-m6A (Synaptic Systems) and other antibodies included in Supplementary information, Data S1 .
Mouse anti mettl3
Manufactured by Abnova
Mouse-anti-METTL3 is a primary antibody that specifically recognizes the METTL3 protein. METTL3 is a methyltransferase that catalyzes the addition of methyl groups to target RNAs, a process known as N6-methyladenosine (m6A) modification. This antibody can be used to detect and study the METTL3 protein in various applications.
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Lab products found in correlation
Pproexhtb, by Thermo Fisher Scientific (1 mentions)
Rabbit anti m6a, by Synaptic Systems (1 mentions)
Pcdna3 ha, by Thermo Fisher Scientific (1 mentions)
P3xflag cmv 14, by Merck Group (1 mentions)
Rabbit anti mettl3, by Abcam (1 mentions)
Pcmv myc, by Thermo Fisher Scientific (1 mentions)
Anti mouse igg alexa fluor 488, by Cell Signaling Technology (1 mentions)
Fv10i confocal microscope, by Olympus (1 mentions)
Dapi containing medium, by Thermo Fisher Scientific (1 mentions)
Rabbit anti mettl14, by Merck Group (1 mentions)
Anti rabbit igg alexa fluor 594, by Abcam (1 mentions)
2 protocols using mouse anti mettl3
1
Cloning and Tagging Human m6A Regulators
2
Visualizing M6A Regulators in 3T3-L1 Adipocytes
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3T3-L1 cells were grown on 0.2% gelatin-coated coverslips and induced to differentiate into adipocytes with standard DMI. At the indicated time point cells were fixed with 95% ethanol, followed by permeabilization with 0.1% Triton X-100 and 0.5% NP-40 on ice. After blocking with 5% nonfat dried milk in TBST, the coverslips were incubated with primary antibody, namely, mouse anti-METTL3 (Abnova), rabbit anti-METTL14 (Sigma), goat anti-WTAP (Santa Cruz), mouse-anti-SC-35 (Abcam), or mouse anti-Smith antigen (Thermo Fisher Scientific), overnight at 4°C. The coverslips then were incubated with fluorescent dye-conjugated secondary antibody, namely, anti-mouse IgG–Alexa Fluor 488 (Cell Signaling Technology), anti-rabbit IgG–Alexa Fluor 594, or anti-goat IgG–Alexa Fluor 647 (Abcam), for 0.5 h at 37°C and mounted with DAPI-containing medium (Life Technologies). Fluorescent images were acquired using an FV10i confocal microscope (Olympus).
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