LSD1 Inhibitor Screening Assay Kit Reference: 700120-96 LSD1 is a histone demethylase whose actions on specific lysine residues alter transcription of chromosomal DNA. It also inhibits the tumor suppressor activity of p53 by demethylating a specific lysine residue. Inhibitors of LSD1 are important tools used to elucidate mechanisms of transcription and cell cycle progression and have therapeutic potential for treating cancer. Cayman’s LSD1 Inhibitor Screening Assay provides a convenient fluorescence-based method for screening LSD1-specific inhibitors. The assay is based on the multistep enzymatic reaction in which LSD1 first produces H2O2 during the demethylation of lysine 4 on a peptide corresponding to the first 21 amino acids of the N-terminal tail of histone H3. In the presence of horseradish peroxidase, H2O2 reacts with ADHP to produce the highly fluorescent compound resorufin that can be analyzed with an excitation wavelength between 530-540 nm and an emission wavelength between 585-595 nm.
Methyltransferase Colorimetric Assay Kit Reference: 700140-96 Methylation of key biological molecules and proteins plays important roles in numerous biological systems, including signal transduction, biosynthesis, protein repair, gene silencing, and chromatin regulation. The SAM dependent MTs use SAM as the enzymatic cofactor. SAM, also known as AdoMet, acts as a donor of a methyl group that is required for the modification of proteins and DNA. Cayman’s MT Colorimetric Assay is a continuous enzyme-coupled assay that can continuously monitor SAM-dependent MT activities. The removal of the methyl group from SAM generates AdoHcy, which is rapidly converted to urate and H2O2 by an enzyme mixture provided in the kit. The rate of production of H2O2 is measured with the colorimetric reagent, 3,5-dichloro-2-hydroxybenzenesulfonic acid, by an increase in absorbance at 500-520 nm. The assay is supplied with AdoHcy as a positive control. The assay can be used with any purified SAM-dependent MT.
Methyltransferase Fluorometric Assay Kit Reference: 700150-96 Cayman’s MT Fluorometric Assay is a continuous enzyme-coupled assay that can continuously monitor SAM-dependent MTs. The removal of the methyl group from SAM generates AdoHcy, which is rapidly converted to urate and H2O2 by an enzyme mixture provided in the kit. The reaction between H2O2 and ADHP (10-acetyl-3,7,-dihydroxyphenoxazine) produces the highly fluorescent compound resorufin, which is analyzed with an excitation wavelength between 530-540 nm and an emission wavelength between 585-595 nm. The assay is supplied with AdoHcy as a positive control. The assay can be used with any purified SAM-dependent MT.