Mononucleosomes, Recombinant Human, Desthiobiotinylated

SKU: 16-0024
Pack Size: 50 µg
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Mononucleosome Recombinant Human Desthiobiotinylated Description: Mononucleosomes assembled from recombinant human histones expressed in E. coli (two each of histones H2A, H2B, H3.1 and H4; accession numbers: H2A-P04908; H2B-O60814; H3.1-P68431; H4-P62805) wrapped with a 147 base pairs of 601 positioning sequence DNA. The nucleosome is the basic subunit of chromatin. The 601 sequence, identified by Lowary and Widom, is a 147-base pair sequence that has high affinity for histone octamers and is useful for nucleosome assembly and contains a 5' desthiobiotin-TEG group. Desthiobiotin has a weaker affinity (Kd = 10-11) for avidin-family proteins than does biotin (Kd = 10-15).

Mononucleosome Recombinant Human Desthiobiotinylated Formulation: Purified recombinant mononucleosomes (50 µg total mass, 27.3 µg protein + 22.7 µg DNA in 42.5 µL) in 10 mM Tris-HCl pH 7.5, 25 mM NaCl, 1 mM EDTA, 2 mM DTT, 20% glycerol. Nucleosome molecular weight = 199,827.86 Da.

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

Mononucleosome Recombinant Human Desthiobiotinylated Application Notes: Mononucleosome, Recombinant Human, Desthiobiotinylated are highly purified and suitable for use as substrates in enzyme screening assays, structural studies, or effector protein binding experiments. The weaker affinity of desthiobiotin for avidin-family proteins allows nucleosomes to be eluted via addition of free biotin. The ability of these nucleosomnes to be recovered intact is useful in many aspects of chromatin biology

Mononucleosome Recombinant Human Desthiobiotinylated References:
Lowary PT and J Widom (1998). J Mol Biol 276: 19-42. Luger K et al (1999). Methods Mol Biol 119: 1-16.

View technical datasheet for this product. 16-0024 Datasheet

16-0024 Protein Gel
Protein Gel Data: Coomassue stained SDS-PAGE of proteins in Mononucleosomes, Human Recombinant, Desthiobiotinylated (1 µg) demonstrates the purity of the histones in the preparation. Sizes of molecular weight markers and positions of the core histones (H2A, H2B, H3, and H4) are indicated.
16-0024 DNA Gel
DNA Gel Data: Mononucleosomes, Recombinant Human, Desthiobiotinylated resolved via native PAGE and stained with ethidium bromide to visualize DNA. Lane 1: Free DNA (200ng). Lane 2: Instact nucleosomes (400ng).
16-0024 Elution Data
Desthiobiotin Elution Data: Mononucleosomes, Recombinant Human, Desthiobiotinylated efficiently elute from strepavidin magentic beads with the addition of 5mM free biotin. Mononucleosomes, Recombinant Human, Biotinylated (EpiCypher Catalog No. 16-0006) do not elute from streptavidin magnetic beads under the same conditions. Lane 1 EpiCypher Catalog No. 18-0005 DNA loading control (100 ng). Lane 2 EpiCypher Catalog No. 16-0006 input (200 ng). Lane 3 1xTBST wash of beads. Lane 4 5mM D(+)-biotin eluate. Lane 5 EpiCypher Catalog No. 18-0024 DNA loading control (100 ng) Lane 6 EpiCypher Catalog No. 16-0024 input (200 ng). Lane 7 1xTBST wash of beads. Lane 8 5mM D(+)-biotin eluate. Nucleosomes remain intact throughout the elution process.
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