A fluorogenic, acetylated peptide substrate for class I HDACs (HDACs 1, 2, and 3) and class 2b HDACs (HDACs 6 and 10). This substrate fluoresces at excitation 350-380 nm, emission at 440-460 nm. If autofluorescence of the inhibitor near this wavelength is an issue, we recommend using Fluorogenic HDAC substrate 2 (green), BPS catalog #50038. For class 2a HDACs (HDACs 4, 5, 7, and 9) or HDACs 8 and 11, we recommend #50040.

5 mM DMSO solution (50 µl)

DMSO solution

Materials :
HDAC Assay buffer (BPS catalog #50031)
HDAC Assay developer (BPS catalog #50030)
HDAC Substrate 3 (BPS catalog #50037)
HDAC1 (0.33 µg/µl) (BPS catalog #50051) or other class1/2b HDAC enzyme

Step 1 :
Add the following reagents to a low binding NUNC black plate (VWR catalog #62408-936):
HDAC Assay Buffer, 38 µl
1 mg/ml BSA, 5 µl
200 µM HDAC substrate 3, 5 µl
HDAC1, 2 µl
Incubate at 37°C for 30 min.

Step 2 :
Stop the reaction by adding 50 µl of HDAC assay developer (undiluted) and incubate the plate at room temperature for 15 min.

Step 3 :
Read samples in a microtiter-plate reading fluorimeter capable of excitation at 350-380 nm and emission at 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

Study enzyme kinetics, and screen small molecule HDAC inhibitors for drug discovery and HTS applications.

1. A. Ito et al. (2001) EMBO J. 20 1331.
2. A. Ito et al. (2002) EMBO J. 21 6236.
3. N.A. Barlev et al. (2001) Mol. Cell 8 1243.

Application Reference(s):
1. Chaperone co-inducer BGP-15 inhibits histone deacetylases and enhances the heat shock response through increased chromatin accessibility (2017) 
2. Design, synthesis, 3D pharmacophore, QSAR, and docking studies of carboxylic acid derivatives as Histone Deacetylase inhibitors and cytotoxic agents (2014)

Protect from direct exposure to light. Avoid freeze/thaw cycles.

Deacetylase

Substrate