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Applications: Great for studying enzyme kinetics and HTS applications.
Contraindications: Green and blue dyes that absorb light in the AlphaScreen® signalemission range (520-620 nm), such as Trypan Blue. Avoid the use of the potent singlet oxygenquenchers such as sodium azide (NaN3) or metal ions (Fe2+, Fe3+, Cu2+, Zn2+ and Ni2+). Thepresence of >1% RPMI 1640 culture medium leads to a signal reduction due to the presence ofexcess biotin and iron in this medium. MEM, which lacks these components, does not affectAlphaScreen® assays.
Description: The SMYD3 Homogeneous Assay Kit is designed to measure SMYD3 activityfor screening and profiling applications. SMYD3 is a histone methyltransferase that promotes diandtrimethylation of lysine4 of histone H3. It also exhibits methylation activity toward a lysineresidue of MAPK/ERK Kinase Kinase 2 (MAP3K2). The key tothe SMYD3 Homogeneous Assay Kit is a highly specific antibody that recognizes methylatedsubstrate. With this kit, only three simple steps on a microtiter plate are required formethyltransferase detection. First, a sample containing SMYD3 enzyme is incubated with thesubstrate for three hours. Next, acceptor beads and primary antibody are added, then donorbeads, followed by reading the Alpha-counts.
Storage Stability: 12 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Supplied As: The SMYD3 Homogeneous Assay Kit comes in a convenient AlphaLISA® format, with GST-tagged SMYD3 substrate, primary antibody, methylation assay buffer, and purified SMYD3 for 384 enzyme reactions.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Dillon, S.C., et al. 2005. Genome Biology 6:227.
2. Silva, F.P., et al. 2008. Oncogene 27:2686.