HeLa

HeLa Mononucleosomes, Purified

$320.00
SKU: 16-0002
Pack Size: 50 μg

Description

Human mononucleosomes 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.

Validation Data*

Figure 1: DNA Gel Data
DNA was purified from HeLa Mononucleosomes 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 Mononucleosomes 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: Histone Methyltransferase Assay Data
HeLa Mononucleosomes were used in a radioactive histone methyltransferase assay with recombinant SET8. Radioactive SAM was incubated with HeLa Mononucleosmes (Nucs, 1.5 μg), recombinant SET8 (SET8, 1 μg), or both (SET8+Nucs). HMTase activity co-purifying with the nucleosomes is virtually undetectable.


*Validation data is representative­­ and may vary slightly between lots. For the current lot's validation data, please see the technical datasheet in the "Documents & Resources" dropdown.

Technical Information

Formulation
Purified HeLa mononucleosomes (50 µg) at a concentration of 0.7 mg/ml (23.1 µg DNA + 26.9 µg protein) in of 71.5 µl of 20 mM HEPES, pH 7.5, 1 mM EDTA. Concentrations may vary between lots.

* Molarity = ~3.27 µM

* MW = ~ 214,000 Da

Storage and Stability
Stable for six months at -80°C from date of receipt. For best results, aliquot and avoid multiple freeze/thaws.

Application Notes

HeLa Mononucleosomes Purified 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 DNA size and accounting for the endogenous post-translational histone modifications.

Note: Despite the high purity of the preparation, a small amount of arginine methyltransferase activity may co-purify 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:
Kuo et al. Mol. Cell (2011). PMID: 22099308
Matthews et al. Nature (2007). PMID: 18033247
Shi et al. Nature (2006). PMID: 16728974

Product References:

Documents & Resources

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