METTL3/METTL14 Complex Chemiluminescent Assay Kit

The METTL3/METTL114 Complex Chemiluminescent Assay Kit is designed to measure the activity of METTL3 (methyltransferase 3, N6-adenosine-methyltransferase complex catalytic subunit) / METTL14 (methyltransferase 14, N6-adenosine) complex activity for screening and profiling applications. The METTL3/METTL14 assay kit comes in a convenient 96-well or 384-well format, with enough recombinant purified METTL3/METTL14 complex, primary and secondary antibodies, S-adenosylmethionine (SAM), pre-coated plate with RNA substrate, blocking buffer and detection reagents for 100 or 400 enzyme reactions.Figure 1: Illustration of the METTL3/METTL114 Complex Chemiluminescent Assay Kit assay principle.Need us to run inhibitor screens or profile your compounds against METTL3/METTL14 complex? Check out ourMethyltransferase Screening Service.

Product Specifications

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.

UniProt

METTL3: Q86U44; METTL14: Q9HCE5

Applications

Study enzyme kinetics and screen small molecule inhibitors for drug discovery and high throughput screening (HTS) applications.

Format

96 ReactionsCatalog #NameAmountStorage100105METTL3/METTL14 Protein Complex, FLAG-Tag, His-Tag*20 μg-80°C5212020 µM S-Adenosylmethionine250 μl-80°C52140Z4Primary Antibody 29100 μl-80°C783684x METTL Assay Buffer3 ml-20°CBlocking buffer 550 ml+4°C0.5 M DTT200 µl-20°C52131HSecondary HRP-labeled antibody 210 μl-80°C79670ELISA ECL substrate A (translucent bottle) 6 mlRoom Temp.ELISA ECL substrate B (brown bottle) 6 mlRoom Temp.White microplate precoated with RNA Substrate1+4°C*The concentration of the protein is lot-specific and will be indicated on the tube.384 ReactionsCatalog #NameAmountStorage100105METTL3/METTL14 Protein Complex, FLAG-Tag, His-Tag*20 μg-80°C5212020 µM S-Adenosylmethionine2 x 250 μl-80°C52140Z4Primary Antibody 292 x 100 μl-80°C783684x METTL Assay Buffer2 x 3 ml-20°CBlocking buffer 52 x 50 ml+4°C0.5 M DTT200 µl-20°C52131HSecondary HRP-labeled antibody 22 x 10 μl-80°C79670ELISA ECL substrate A (translucent bottle) 2 x 6 mlRoom Temp.ELISA ECL substrate B (brown bottle) 2 x 6 mlRoom Temp.White microplate precoated with RNA Substrate1+4°C*The concentration of the protein is lot-specific and will be indicated on the tube.

Shipping Conditions

-80°C

Storage Conditions

This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

Notes

Troubleshooting GuideVisitbpsbioscience.com/assay-kits-faqfor detailed troubleshooting instructions. For all further questions, please email[email protected]