Fluorogenic HDAC class 2a substrate

Catalog #: 50040
Size: 250 nmole
A fluorogenic, acetylated peptide substrate for class2a HDACs (HDACs 4, 5, 7, and 9). It also works well for HDACs 8 and 11. 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 class 2a substrate 2 (green), BPS catalog #50042. For HDACs 1-3 ,6, and 10, we recommend substrate #50037.
Supplied As:
5 mM DMSO solution (50 µl)
DMSO solution
Instructions for use:
Materials :
HDAC Assay buffer (BPS catalog #50031)
HDAC Assay developer (BPS catalog #50030)
HDAC Class 2A Substrate (BPS catalog #50040)
HDAC4 (0.2 ng/µl) (BPS catalog #50004) or other class2a HDAC enzyme

Step 1 :
Add the following reagents to a low binding NUNC black plate (VWR catalog #62408-936):
HDAC Assay Buffer, 35 µl
1 mg/ml BSA, 5 µl
200 µM HDAC 2A substrate, 5 µl
HDAC4, 5 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.
Storage / Stability:

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

Study enzyme kinetics, and screen small molecule inhibitors of HDAC class2a for drug discovery and HTS applications.
Protect from direct exposure to light. Avoid freeze/thaw cycles.
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