Fluorogenic HDAC Substrate 2 (Green fluorescence)

Catalog #: 50038
Size: 250 nmole
$230
Description:
A fluorogenic, acetylated peptide substrate for HDACs. It is a suitable substrate for class I HDACs (HDACs 1, 2, and 3) and class 2b HDACs (HDACs 6 and 10). This substrate exhibits longer excitation and emission wavelengths (excitation 485 nm, emission 528 nm) than our regular class 1/2b HDAC substrate, #50037, which fluoresces at excitation 350-380 nm, emission at 440-460 nm. This substrate should be used when the inhibitor fluoresces near 450 nm. If autofluorescence is not an issue, we recommend HDAC substrate 3, #50037. For autofluorescent samples using HDACs 4, 5, 7, 8, 9, and 11, we recommend substrate #50042.
Supplied As:
5 mM DMSO solution (50 µl)
Format:
DMSO solution
Instructions for use:
Materials :
HDAC Assay buffer (BPS catalog #50031)
HDAC Assay developer (BPS catalog #50030)
HDAC Substrate 2 (green) (catalog #50038)
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 2, 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 485 nm and emission at 528 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

Application(s):
Study enzyme kinetics, and screen small molecule HDAC inhibitors for drug discovery and HTS applications.
Reference(s):

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.

Product Specific Reference(s):
1. Day, JA., et al.,  J Med Chem. 2013 Oct 24;56(20):7997-8007. 

Application Reference(s):

1. The first report of direct inhibitors that target the C-terminal MEEVD region on heat shock protein 90 (2016)
Warning(s):
Protect from direct exposure to light. Avoid freeze/thaw cycles.
Scientific Category:
Deacetylase
Product Type:
Substrate
Data shown is lot-specific. Contact us for specific information on other lots.

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Product Datasheets

50038_Lot130903.pdf