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Application: Study enzyme kinetics and screen small molecule inhibitors for drug discovery and high throughput screening (HTS) applications.
Background: METTL3 (methyltransferase 3, N6-adenosine-methyltransferase complex catalytic subunit)/METTL4 (methyltransferase 4, N6-adenosine) and METTL14 (methyltransferase 14, N6-adenosine) form the core of a complex involved in N6-methyladenosine addition to RNA, one of the most common mRNA modifications, and belong to the group known as "writer" proteins. METTL3 expression is altered in cancer via different processes. For instance, in pancreatic cancer cigarette smoke condensate influences METTL3 promoter hypomethylation and increases its expression. This complex can act as a tumor suppressor or activator and promote chemoresistance. It has been linked to acute myeloid leukemia (AML), liver, lung, bladder cancer and others. Interestingly METTL3 can activate certain oncogenes by simply recruiting eIF3 (eukaryotic translation initiation factor 3), a process that does not require its catalytic activity. Its role in cancer has made it an attractive target in oncology. STM2457, a highly selective potent METT3/METTL14 inhibitor, resulted in AML cells undergoing differentiation and apoptosis. A deeper understanding of the role of this complex in cancer and development of new therapeutics around it will benefit the cancer therapy field.
Contraindications: DMSO >1%, strong acids or bases, ionic detergents, high salt.
Description: The METTL3/METTL14 Complex Chemiluminescent Assay Kit is designed to measure METTL3/METTL14 complex activity for screening and profiling applications. The key to the METTL3/METTL14 Complex Chemiluminescent Activity Assay Kit is a highly specific antibody that recognizes N6-methylated adenosine. With this kit, only three simple steps are required for methyltransferase detection. First, S-adenosylmethionine is incubated with a sample containing assay buffer and methyltransferase enzyme. Next, primary antibody is added. Finally, the plate is treated with an HRP-labeled secondary antibody followed by addition of the HRP substrate to produce chemiluminescence that can then be measured using a chemiluminescence reader.
Format: 96 Reactions
Storage Stability: 6 months from date of receipt when stored as directed.
Supplied As: The METTL3/METTL14 Complex Chemiluminescent Assay Kit comes in a convenient format, with 96-well plate precoated with a specific substrate, the antibody against methylated adenosine, a secondary HRP-labeled antibody, S-adenosylmethionine, methylt
Uniprot: METTL3: Q86U44; METTL14: Q9HCE5
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Liu, J., et al. Nat Chem Biol. 2014; 10(2): 93-95.
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