Fluorogenic Rat HDAC6 Assay Kit
The rat histone deacetylase 6 (HDAC6) Assay Kit is a 96-well fluorogenic assay designed to measure rat HDAC6 enzymatic activity for screening and profiling applications. In addition, the kit includes a potent HDAC inhibitor, Trichostatin A (TSA), for use as control.
Figure 1: Illustration of the principle behind the Fluorogenic Rat HDAC Assay Kit.
The Fluorogenic Rat HDAC6 Assay Kit is based on a unique fluorogenic substrate and developer combination. A fluorescent dye is conjugated to a peptide containing acetyl-lysine. The fluorescence of the conjugated dye is quenched by the peptide. After treatment of the peptide with an HDAC, a developer solution specific for non-acetylated lysine is added to the reaction. If the acetyl group has been removed from the lysine, the developer releases the dye, allowing for fluorescence. In this assay, fluorescence directly correlates with HDAC activity.
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- 1 mg/ml of bovine serum albumin (BSA) dissolved in distilled water
- Fluorescent microplate reader capable of reading λexc/λem=350-380 nm/440-460 nm
- Adjustable micropipettor and sterile tips
| Catalog # | Name | Amount | Storage |
| 101332 | HDAC6, GST-His-Tag (Rat)* | 9 µg | -80°C |
| 50037 | Fluorogenic HDAC substrate 3 (5 mM) | 25 µl | -80°C |
| 50030 | 2x HDAC Developer (contains Trichostatin A) (2 µM) |
6 ml | -80°C |
| Trichostatin A in DMSO (200 µM)** |
100 µl | -20°C | |
| 50031 | HDAC Assay Buffer | 10 ml | -20°C |
| 79685 | Black 96-well plate | 1 | Room Temp |
*The concentration of the protein is lot-specific and will be indicated on the tube.
**HDAC6 inhibitor Trichostatin A is provided as an internal control for HDAC6 activity.
Histone Deacetylases (HDACs) catalyze the hydrolysis of an acetyl group from acetyl-lysine present in histones. Histone acetylation/deacetylation status tightly regulates transcription. HDAC6 plays a role in stress granule formation, cell motility, and leptin sensitivity. HDAC6 interacts with heat-shock protein 90 (HSP90) and with ubiquitinated proteins and has been linked to Alzheimer’s disease and tumor growth and metastasis. The use of selective inhibitors for HDAC6 offers potential benefits for neurodegenerative diseases and cancer treatment.
Hey J. et al., 2023, HDAC6 score: to treat or not to treat? Nature Cancer. 4: 156-158
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