HDAC assay buffer is an essential reagent for signal generation in HDAC assays performed with any of BPS Bioscience's HDAC fluorimetric substrates or assay kits. An appropriate fluorimetric substrate, comprising an acetylated lysine side chain, is first incubated with a sample containing HDAC activity (nuclear or cellular extract, purified enzyme, bead-bound immunocomplex, etc.) in HDAC assay buffer. Deacetylation of the substrate sensitizes it so that, in a second step, treatment with the HDAC assay developer produces a fluorophore. Trichostatin A is included as an ‘inhibitor stop’ for class I and II HDACs.

Tris-buffered solution

Aqueous buffer solution

In a 50 µL reaction, assay HDAC activity using HDAC (enzyme, lysate, etc.), substrate, HDAC assay buffer, and test inhibitor. Incubate microtiter plate at 37°C for 30 minutes. Add an equal volume (50 µL) of HDAC Assay Developer to each well. Incubate the plate at room temperature for 15 minutes. Read sample in a microtiter-plate reading fluorimeter capable of excitation at a wavelength in the range of 350-380 nm and detection of emitted light in the range of 440-460 nm. “Blank” value (no enzyme negative control) is subtracted from all other values.

Stable at least 12 months from date of receipt, when stored as directed

Suitable for use with cell extracts, nuclear extracts, immunoprecipitates, and purified HDAC enzymes.

Application Reference:
Design, synthesis, 3D pharmacophore, QSAR, and docking studies of carboxylic acid derivatives as Histone Deacetylase inhibitors and cytotoxic agents (2014)

Do not dilute! Avoid freeze/thaw cycles

Deacetylase

Substrate

HDAC Assay Developer #50030 6 mL
SIRT Assay Developer #50089 6 mL
SIRT1 (Sir2) Assay Kit #50081 100 rxns.
HDAC-1 Active Enzyme #50051 50 μg
HDAC-2 Active Enzyme #50002 50 μg
HDAC-3 Active Enzyme #50003 50 μg
HDAC-4 Active Enzyme #50004 10 μg
HDAC-5 Active Enzyme #50005 10 μg
HDAC-6 Active Enzyme #50006 50 μg
HDAC-7 Active Enzyme #50007 10 μg