SIRT6 Direct Fluorescent Screening Assay Kit

Cayman Chemical Item Number 700290

SIRT6 Direct Fluorescent Screening Assay Kit

Description

SIRT6 is a nuclear protein that has both ADP-ribosyl transferase and deacetylase activities. SIRT6 associates specifically with telomeres and functions at chromatin to decrease NF-κB signaling. Cayman’s SIRT6 Direct Fluorescent Screening Assay provides a convenient fluorescence-based method for screening SIRT6 inhibitors or activators. The procedure requires only two easy steps, both performed in the same microplate. In the first step, the substrate, which comprises the p53 sequence Arg-His-Lys-Lys(ε-acetyl)-AMC, is incubated with human recombinant SIRT6 along with NAD⁺. Deacetylation sensitizes the substrate such that treatment with the developer in the second step releases a fluorescent product having excitation and emission at 350-360 nm and 450-465 nm, respectively.

Stability

1 year

Storage

-80°C

Shipping

Dry ice in continental US; may vary elsewhere

Background Reading

Tanny, J.C., and Moazed, D. Coupling of histone deacetylation to NAD breakdown by the yeast silencing protein Sir2: Evidence for acetyl transfer from substrate to an NAD breakdown product. Proc Natl Acad Sci USA 98(2) 415-420 (2001).

Saunders, L.R., and Verdin, E. Sirtuins: critical regulators at the crossroads between cancer and aging. Oncogene 26 5489-5504 (2007).

Frye, R.A. Phylogenetic classification of prokaryotic and eukaryotic Sir2-like proteins. Biochem Biophys Res Commun 273 793-798 (2000).

Michishita, E., McCord, R.A., Berber, E., et al. SIRT6 is a histone H3 lysine 9 deacetylase that modulates telomeric chromatin. Nature 452 492-496 (2008).

Cheung, W.L., Briggs, D.B., and Allis, C.D. Acetylation and chromosomal functions. Curr Opin Cell Biol 12 326-333 (2000 Jan 1).

Tanner, K.G., Landry, J., Sternglanz, R., et al. Silent information regulator 2 family of NAD-dependent histone/protein deacetylases generates a unique product, 1-O-acetyl-ADP-ribose. Proc Natl Acad Sci USA 97(26) 14178-14182 (2000).

Grunstein, M. Histone acetylation in chromatin structure and transcription. Nature 389 349-352 (1997 Sep 25).

Imai, S., Armstrong, C.M., Kaeberlein, M., et al. Transcriptional silencing and longevity protein Sir2 is an NAD-dependent histone deacetylase. Nature 403 795-800 (2000).

Kawahara, T.L.A., Michishita, E., Adler, A.S., et al. SIRT6 links histone H3 lysine 9 deacetylation to NF-kB-dependent gene expression and organismal life span. Cell 136 62-74 (2009).

Strahl, B.D., and Allis, D. The language of covalent histone modifications. Nature 403 41-45 (2000 Jan 6).

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700290-96WELLS

Half Volume 96-Well Solid Plate (white)