SensoLyte® HDACs and HATs Assay Kits – Highly Sensitive


AnaSpec, EGT Group is pleased to offer a wide selection of highly sensitive HDAC (HDAC, SIRT1 & SIRT2) and HAT (CAF & p300) SensoLyte® assay kits. These assays are fluorimetric, some of which are FRET (fluorescence resonance energy transfer)-based.

Histones are the chief protein components of chromatin. They act as spools around which DNA winds. Covalent modification of histone proteins through acetylation and deacetylation affects chromatin structure and regulates gene expression. Histone hyperacetylation is well correlated with increased transcription, whereas hypoacetylation correlates with transcriptional repression.1 Histone deacetylases (HDACs), which catalyze the removal of acetyl groups from a ε-N-acetyl lysine amino acid on a histone, act as transcriptional repressors of genes. Histone deacetylases have been grouped into three classes. Class I (HDAC 1, 2, 3, 8) and Class II (HDAC4, 5, 6, 7, and 9) are zinc-containing hydrolase enzymes.2,3 The third class of deacetylases consists of the members of the sirtuin family of enzymes (Sir 1 to 7).4 Inhibitors of HDAC classes I and II are being studied as a treatment for cancer and neurodegenerative diseases such as Huntington's and Alzheimer's diseases.5-7 The Sirtuin 1 (class III) enzyme represents a target for treatment of age-related diseases and type II diabetes.8,9

Histone acetyltransferases (HATs) regulate the acetylation of histones and non-histone proteins.10, 11 The acetylation of the ε-amino groups of lysine residues present at histone tails correlates largely with transcriptional activation, and is also involved in DNA replication, DNA repair and protein–protein interactions.12


Product

Catalog #

SensoLyte® 520 HDAC Activity Assay Kit *Fluorimetric*

72084

SensoLyte® 440 HDAC Activity Assay Kit *Fluorimetric*

72083

SensoLyte® 520 FRET SIRT1 Assay Kit *Fluorimetric*

72155

SensoLyte® Green SIRT1 Assay Kit *Fluorimetric*

72156

SensoLyte® 520 FRET SIRT2 Assay Kit *Fluorimetric*

72189

SensoLyte® Green SIRT2 Assay Kit *Fluorimetric*

72188

SensoLyte® HAT (p300) Assay Kit *Fluorimetric*

72172

SensoLyte® HAT (pCAF) Assay Kit *Fluorimetric*

72173

SensoLyte® LSD1 Assay Kit *Fluorimetric*

72131

SensoLyte® 520 Deubiquitination Assay Kit *Fluorimetric*

72203

SensoLyte® 440 Deubiquitination Assay Kit *Fluorimetric*

72204



Figure 1. HDAC substrates were incubated with HeLa nuclear extracts, followed by incubation of a Trichostatin A-containing developer. The substrate in SensoLyte® 520 provides higher sensitivity and better linear range.


Figure 2. The SensoLyte® Green and 520 FRET SIRT1 substrates provide better signal/background ratio than the AMC-containing substrate.


Figure 3. AMC, Green and FRET SIRT2 substrates were incubated with sirtuin 2 enzyme. The substrates in the SensoLyte® 520 FRET and Green SIRT assay kits provide superior signal/background ratio.


Figure 4. Inhibitor studies. The calculated IC50 for sirtuin inhibitor Ro-31-8220 was about 2 mM for both enzymes in the fluorogenic and FRET assays. SIRT2 was also tested with suramin. The calculated IC50 were 8.7 mM and 19.7 mM for fluorogenic and FRET substrates, respectively.


Related Products

Other Assay Kits

SensoLyte® Histone H3 (1 - 21) Sampler Kit *Biotinylated*

SensoLyte® Methylated Histone H3 (1 - 21) Sampler Kit *Biotinylated*

Peptides

Histone H1 Peptides

Histone H2A Peptides

Histone H2B Peptides

Histone H3 Peptides

Histone H4 Peptides

Histone Related Substrates

Antibodies

Anti-Histone H2B (acetyl K5), polyclonal

Anti-Histone H1

Anti-H4 & H2A (paired1)

Anti-LSD1

Anti-HDAC-1 (IN), Z-Fish™

View the following posters that had been presented at conferences:

Development of Histone Deacetylase Activity Assay Using a Novel Long Wavelength Fluorogenic Substrate

Design of Novel Fluorogenic and FRET Substrates for the Detection of Sirtuin Activity



References

1. Sterner, DE. et al. Microbiol. Mol. Biol. Rev. 64, 435 (2000).
2. De Ruijter, AJ. et al. Biochem. J. 370, 737 (2003).
3. Verdin, E. et al. Trends Genet. 19, 286 (2003).
4. Blander, G. and L. Guarente Annu. Rev. Biochem. 73, 417 (2004).
5. Huang, L. J. Cell Physiol. 209, 611 (2006).
6. Dompierre, JP. et al. J. Neurosci. 13, 3571 (2007).
7. Langley, B. Curr. Drug Targets CNS Neurol. Disord. 4, 41 (2005).
8. Trapp, J. and Jung, M. Curr. Drug Targets 7, 1553 (2006).
9. Porcu, M., and Chiarugi, A. Trends Pharmacol Sci. 26, 94 (2005).
10. Roth, SY. et al. Annu Rev Biochem. 70, 81 (2001).
11. Glozak, MA. et al. Gene. 363, 15 (2005).
12. Turner, BM. Bioessays 22, 836 (2000).