Nucleosome Preparation Kit

by Active Motif
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Description

Active Motif’s Nucleosome Preparation Kit is designed to prepare mono- and oligonucleosomes starting from cells. Nucleosomes comprise the smallest subunit of chromatin and consist of 147 bp of DNA wrapped around an octamer of core histone proteins (H2A, H2B, H3 and H4). Each nucleosome is separated by a linker of DNA. The Nucleosome Preparation Kit uses an enzymatic cocktail to digest the linker DNA. By adjusting the digestion time, it is possible to control the amount of mononucleosomes or oligonucleosomes extracted. Because isolated nucleosomes are more physiologically relevant than histones alone or synthetic peptides, they are ideally suited for use as substrates in the analysis of histone post-translational modifications, enzyme kinetics or inhibitor screening studies.

Nucleosome Preparation Kit preserves intact nucleosomes.
Gel shift showing preservation of intact nucleosomes.

Nucleosome Advantages:

  • More physiologically relevant substrates than synthetic peptides
  • Useful for the analysis of histone post-translational modifications, enzyme kinetics or inhibitor screening studies
  • Easily adjust the levels of mononucleosomes or oligonucleosomes isolated by adjusting digestion times
 
Name Format Cat No. Price  
Nucleosome Preparation Kit 20 rxns 53504 $265 Buy Now

Easily prepare mononucleosomes or oligonucleosomes from your samples

The ability to regulate the amount of mononucleosomes or oligonucleosomes isolated with the Nucleosome Preparation Kit based on digestion time with the Enzymatic Shearing Cocktail is shown in Figure 1. A comparison of isolated nucleosomes with purified nucleosome DNA in Figure 2 illustrates the gel shift observed in the molecular weight of the nucleosome sample, thereby confirming the nucleosome is intact with DNA wrapped around the histone proteins.

Efficiency of nucleosome digestion into mononucleosomes and oligonucleosomes
Figure 1: DNA gel analysis of nucleosome digestion times.

Nucleosomes were prepared from HeLa cells and digested with the Enzymatic Shearing Cocktail for 5, 10 and 15 minutes. Following digestion, the reactions were stopped with the addition of ice-cold EDTA. The nucleosomes were then subjected to DNA clean up to assess the digestion efficiency. Samples were separated by electrophoresis through a 1.5 % agarose gel

Lane 1: 100 to 1000 bp DNA ladder.
Lane 2: H

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