Functional Assays

SIRT Assay Developer is an essential reagent for signal generation in sirtuin assays performed with any of BPS Bioscience's SIRT substrate or fluorimetric assay kits. An appropriate fluorimetric substrate, comprising an acetylated lysine side chain, is first incubated with a sample containing SIRT activity (nuclear or cellular extract, purified enzyme, bead-bound immunocomplex, etc.). Deacetylation of the substrate sensitizes it so that, in a second step, treatment with the SIRT assay developer produces a fluorophore. Nicotinamide is included as an 'inhibitor stop' for Sirtuins.

Histone H3 peptide methylated at Lysine 4, substrate for LSD1

The LSD1 Homogeneous Assay Kit is designed to measure activity of lysine-specific demethylase (LSD1) for screening and profiling applications. The key to the LSD1 Homogeneous Assay Kit is a highly specific antibody that recognizes demethylated substrate. With this kit, only three simple steps on a microtiter plate are required for demethylase detection. First, a sample containing LSD1 enzyme is incubated with the biotinylated substrate. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts.

The LSD1 Chemiluminescent Assay Kit is designed to directly measure activity of human lysine-specific demethylase (LSD1) for screening and profiling applications. The key to the LSD1 Chemiluminescent Assay Kit is a highly specific antibody that recognizes demethylated substrate. With this kit, only three simple steps on a microtiter plate are required for methyltransferase detection. First, a sample containing LSD1 enzyme is incubated with a sample containing assay buffer. 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.

A fluorogenic, succinylated peptide substrate for Sirtuin 5. This substrate should be used in conjunction with SIRT Developer, #50089. Optimal excitation wavelength 350-380 nm, emission wavelength 440-460 nm.

The FBXL10 Chemiluminescent Assay Kit is designed to measure FBXL10 activity for screening and profiling applications. FBXL10, also known as KDM2B, JHDM1B, and CXXC2, is a histone lysine demethylase that exhibits demethylation activity toward trimethyllysine 4 (H3K4me3) and dimethyl-lysine 36 (H3K36me2) on histone H3. The key to the FBXL10 Chemiluminescent Assay Kit is a highly specific antibody that recognizes demethylated substrate. With this kit, only three simple steps on a microtiter plate are required for detection of demethylase activity. First, FBXL10 enzyme is incubated with the methylated H3 peptide for one hour. Next, primary antibody is added. Finally, the plate is treated with an HRP-labeled secondary antibody followed by the addition of the HRP substrate to produce chemiluminescence that can be measured using a chemiluminescence reader.

Histone H3 peptide di-methylated at Lysine 4, substrate for LSD1

The BCL-2 TR-FRET Assay Kit is designed to measure the inhibition of BCL-2 binding to its ligand in a homogeneous 384 reaction format. This FRET-based assay requires no time-consuming washing steps, making it especially suitable for high throughput screening applications. The assay procedure is straightforward and simple; a sample containing terbiumlabeled donor, dye-labeled acceptor, BCL-2, peptide ligand, and an inhibitor is incubated for 120 minutes. Then, the fluorescence intensity is measured using a fluorescence reader.

The BCL-XL TR-FRET Assay Kit is designed to measure the inhibition of BCLXL binding to its ligand in a homogeneous 384 reaction format. This FRET-based assay requires no time-consuming washing steps, making it especially suitable for high throughput screening applications. The assay procedure is straightforward and simple; a sample containing terbium-labeled donor, dye-labeled acceptor, BCL-XL, peptide ligand, and an inhibitor is incubated for 120 minutes. Then, the fluorescence intensity is measured using a fluorescence reader.