HDAC Fluorogenic Assay Kit (Green)
The HDAC Fluorogenic Assay Kit (Green) is designed to measure HDAC (histone deacetylase) class I (HDAC1, 2, and 3) and Class II (HDAC6) activity for screening and profiling applications. The assay kit comes in a convenient 96-well format, with enough fluorogenic substrate, HDAC Developer and assay buffer for 100 enzyme reactions. This kit also contains recombinant purified HDAC2 and the inhibitor Trichostatin as control.
Figure 1: Illustration of the mechanism behind the HDAC Fluorogenic Assay Kit (Green).
The fluorescence from dye molecules is quenched when bound to the peptide substrate. HDAC catalyzes the hydrolysis of the acetyl group from the lysine. Upon incubation with a developer solution specific for non-acetylated lysines, the dye is released and able to fluoresce (λex=485 nm; λem=528 nm). Fluorescence is thus proportional to HDAC activity.
This product has been cited 28 times.
- 1 mg/ml BSA (bovine serum albumin) solution in distilled water
- Fluorimeter capable of excitation at λ=485 nm and detection at λ=528 nm
- Adjustable micropipettor and sterile tips
- Orbital shaker
| Catalog # | Name | Amount | Storage |
| 50002 | HDAC2, His-Tag* | 5 μg | -80°C |
| 50038 | 5 mM Fluorogenic HDAC Substrate 2 | 25 μl | -80°C |
| 50030 | 2x HDAC Developer (contains 2 μM Trichostatin A) | 6 ml | -80°C |
| 200 μM Trichostatin A | 100 μl | -20°C | |
| 50031 | HDAC Assay Buffer | 10 ml | -20°C |
| 79685 | Black, low binding microtiter plate | 1 | Room Temp |
*The concentration of the protein is lot-specific and will be indicated on the tube.
HDACs, or histone deacetylases, are a class of proteins involved in lysine deacetylation (removal of acetyl groups). Lysine acetylation/deacetylation is a dynamic process involved in the regulation of a variety of cellular functions, similarly to phosphorylation/dephosphorylation. Acetylation is performed by histone acetyltransferases. There are four classes of HDACs, classified based on sequence homology and domain organization. Class I include HDAC1, 2, 3, and 8, class III includes sirtuin proteins and class IV has only one member, HDAC11. Class II is subdivided into two subclasses, with Class IIA including HDAC4, 5, 7, and 9, and Class IIB including HDAC6 and 10. Mutations in HDACs can lead to pathologies. For instance, dysfunction of HDAC1 can result in cancer, via the deregulation of genes involved in cell proliferation and survival. Several HDAC inhibitors have been approved for the treatment of HDAC-linked diseases, but most are non-selective. The development of new inhibitors specifically targeting HDAC may open newer avenues for cancer and other HDAC-linked diseases in the endothelium.
Ito A., et al., 2001 EMBO J. 20: 1331.
Barlev N.A, et al., 2001 Mol. Cell 8: 1243.
Ito A., et al., 2002 EMBO J. 21: 6236.
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