MLL1 SAM-Screener™ Assay Kit Reference: 600580-1920 Most histone lysine methyltransferases contain a conserved domain (SET) that utilizes S-adenosyl-L-methionine (SAM or AdoMet) as a co-factor to catalyze the methylation of the epsilon amino group of lysine. Mixed-lineage leukemia (MLL1) is a member of the trithorax group (trxG)/Set1-like family of gene activators that contains histone methyltransferase activity specific for lysine 4 of histone H3. This methylation plays an important role in gene activation at various developmentally regulated loci, such as the Hox gene loci.This fluorescence polarization assay is based upon a proprietary small molecule fluorescent probe that binds to the SAM binding pocket in MLL1. Binding of the small molecule probe to MLL1 induces an increase in fluorescence polarization. Binding of the probe can be competed with the endogenous cofactor SAM or by the inhibitor sinefungin, but is unaffected by the histone H3 peptide substrate. The MLL1 SAM-Screener Assay is robust (Z' >0.6) and exhibits a greater than 100 mP shift over a range of 0-500 nM MLL1. The assay is suitable for high-throughput screening in the provided 384-well plate or can be scaled to higher density plate formats (e.g., 1,536-well) if desired.
MLL1 SAM-Screener™ Assay Kit Reference: 600580-384 Most histone lysine methyltransferases contain a conserved domain (SET) that utilizes S-adenosyl-L-methionine (SAM or AdoMet) as a co-factor to catalyze the methylation of the epsilon amino group of lysine. Mixed-lineage leukemia (MLL1) is a member of the trithorax group (trxG)/Set1-like family of gene activators that contains histone methyltransferase activity specific for lysine 4 of histone H3. This methylation plays an important role in gene activation at various developmentally regulated loci, such as the Hox gene loci.This fluorescence polarization assay is based upon a proprietary small molecule fluorescent probe that binds to the SAM binding pocket in MLL1. Binding of the small molecule probe to MLL1 induces an increase in fluorescence polarization. Binding of the probe can be competed with the endogenous cofactor SAM or by the inhibitor sinefungin, but is unaffected by the histone H3 peptide substrate. The MLL1 SAM-Screener Assay is robust (Z' >0.6) and exhibits a greater than 100 mP shift over a range of 0-500 nM MLL1. The assay is suitable for high-throughput screening in the provided 384-well plate or can be scaled to higher density plate formats (e.g., 1,536-well) if desired.
Nrf2 Transcription Factor Assay Kit Reference: 600590-96 Nuclear factor E2-related factor 2 (Nrf2) is a transcription factor that plays a key role in maintaining redox homeostasis via its interaction with a cysteine-rich protein Kelch-like ECH-associated protein 1 (Keap1). In resting cells, Nrf2 and Keap1 form a tight complex, which is targeted for degradation by proteasomes. Under oxidative stress, Nrf2 is released from the Nrf2/Keap1 complex and translocates to the nucleus where it is able to induce the expression of a battery of genes encoding diverse cytoprotective proteins, including antioxidative enzymes, anti-inflammatory mediators, and proteasomes. Cayman’s Nrf2 Transcription Factor Assay is a non-radioactive, colorimetric method for detecting specific transcription factor DNA binding activity in nuclear extracts. A specific double stranded DNA (dsDNA) sequence containing the Nrf2 response element is immobilized onto the wells of a 96-well plate. Nrf2 contained in nuclear extract samples will bind specifically to the Nrf2 response element and can be detected by addition of a specific Nrf2 antibody and a secondary antibody conjugated to HRP.