HeLa

HeLa Polynucleosomes, Purified

$320.00
SKU: 16-0003
Pack Size: 50 μg

Description

Human Polynucleosomes purified from HeLa cells. The nucleosome is the basic subunit of chromatin consisting of the histone octamer (two each of the four core histones, H2A, H2B, H3 and H4) wrapped by 147 base pairs of DNA.

HeLa Polynucleosomes are predominantly trimers, with some dimers and tetramers.

Validation Data

Figure 1: DNA Gel Data
DNA was purified from HeLa Poly-nucleosomes, Purified and run on an agarose gel to show the size of nucleosomal DNA compared to molecular weight markers (base pairs). >

Figure 2: Protein Gel Data
Coomassie stained PAGE gel of proteins in HeLa Polynucleosomes, Purified to demonstrate the purity of the histones in the preparations. Sizes of molecular weight markers (kDa) and position of the core histones are indicated.

Figure 3: HMTase Assay Data
HeLa Polynucleosomes were used in a radioactive histone methyltransferase assay with recombinant SET8. Radioactive SAM was incubated with HeLa Polynucleosmes (Nucs, 1.5 μg), recombinant SET8 (SET8, 1 μg), or both (SET8+Nucs). HMTase activity co-purifying with the nucleosomes is virtually undetectable.

Technical Information

Formulation

Purified HeLa Polynucleosomes (50 µg) at a concentration of 0.8 mg/ml (24.8 µg DNA + 25.2 µg protein) in of 62.5 µl of 20 mM HEPES, pH 7.5, 1 mM EDTA.

*Molarity = ~3.5µM. * MW = ~ 230,000 Da

Concentration may differ from previous lots.

Storage and Stability
Stable for six months at -80°C from date of receipt. It is recommended to aliquot samples and do not refreeze.

Application Notes

HeLa Polynucleosomes are suitable for use in enzyme assays such as acetylation or methylation, chromatin binding assays, or for use as a positive control in Western blotting. Use 1-2 µg per reaction.

*-Molarity and molecular weight are estimates based on average DNA size (including linker) and accounting for the endogenous post-translational histone modifications.

NB. Despite the high purity of the preparation, a small amount of arginine methyltransferase activity co-purifies with the nucleosomes. If you are studying weak or low turnover HMTases, you can use AMI-1 to inhibit the endogenous RMTase activity.

References

Background References:
Shi et al. Nature (2006) PMID: 16728974
Matthews et al. Nature (2007) PMID: 18033247
Kuo et al. Mol. Cell (2011) PMID: 22099308

Product References:
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