Nucleosome Preparation Kit
easily isolate mononucleosomes and oligonucleosomes
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.
Gel shift showing preservation of intact nucleosomes.
- 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
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.
Figure 1: DNA gel analysis of nucleosome digestion times.
Figure 2: Gel shift observed with intact nucleosomes as compared to purified nucleosome DNA.
Contents & Storage
Please note that the Nucleosome Preparation Kit is shipped on dry ice and contains reagents with multiple storage temperatures inside. Please store each component at the temperature indicated below. All reagents are guaranteed stable for 6 months from date of receipt when stored properly. This kit includes enough reagents for 20 preparations from 10 cm plates and 5 sample optimizations.
- 10X PBS; Store at 4°C to -20°C
- 100 mM PMSF; Store at -20°C
- Protease Inhibitor Cocktail (PIC); Store at -20°C
- Lysis Buffer; Store at -20°C
- Digestion Buffer; Store at 4°C to -20°C
- Enzymatic Shearing Cocktail; Store at -20°C
- 0.5 M EDTA; Store at 4°C to -20°C
- RNase A (10 µg/µl); Store at -20°C
- Proteinase K (10 µg/µl); Store at -20°C
- 5 M NaCl; Store at -20°C
- 3 M Sodium Acetate; Store at RT