Recombinant Human Nucleosome, H2A-Ubiquityl Protein, CF

Catalog #: 16-0020 Datasheet / COA / SDS

Discontinued Product

16-0020 has been discontinued.
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Recombinant Human Nucleosome, H2A-Ubiquityl Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.
Activity
Concentrations for in vitro assays will depend on experimental conditions and detection methods.
Source
E. coli-derived human Nucleosome protein
Predicted Molecular Mass
227 kDa, average

Product Datasheets

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16-0020

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

16-0020

Formulation X mg/ml in 25 mM Tris pH 7.5, 150 mM NaCl, 2 mM DTT, 20% (v/v) Glycerol
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
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Background: Nucleosome

Within the context of chromatin, post-translational modifications (PTM's) of histones play important roles in nearly every biologically occurring DNA-related process.  Ubiquitination is one such histone PTM, and while the modification was discovered more than three decades ago, its functions are less well understood than other histone modifications such as methylation and acetylation.  Polycomb Repressive Complex 1 (PRC1) is an evolutionarily conserved, multi-subunit protein that is essential for maintaining epigenetic repression of key developmental loci in embryonic stem cells (ESCs).  PRC1 proteins BMI1, RNF2 and others function together as a Ubiquitin E3 ligase that specifically mono-ubiquitinates histone H2A (H2AK119u1) which effects chromatin remodeling in a heritably transmissible manner.  BMI1 activity has also been reported as necessary for constitutive heterochromatin formation in mammalian somatic cells, again demonstrating that ubiquitination of histones is important in maintaining genomic integrity and transcriptional control in eukaryotes.

Produced and offered in collaboration with EpiCypher™, this mononucelosome is assembled from recombinant human histones expressed in E.coli (two each of histones H2A, H2B, H3 and H4.  Accession numbers P04908, O60814, P68431 and P62805, respectively) wrapped by 147 base pairs of 601 positioning sequence DNA.  There is a 5' biotin-TEG group on the DNA.  Approximately 50% of the nucleosomes are mono-ubiquitinated on histone H2A lysine 118, while the other 50% are mono-ubiquitinated on both histone H2A lysine 118 and histone H2A lysine 119 (multi-mono-ubiquitination).

References
  1. Abdouh M. et al (2016) J. Biol. Chem.  291: 182
  2. Cao J. & Yan Q. et al. (2012) Front. Oncol.  2: 26
  3. Endoh M. et al. (2012) PLoS doi.org/10.1371/journal.pgen.1002774
  4. Lowary P.T. & Widom J (1998) J. Mol. Biol.  276: 19
  5. Luger K. et al. (1999) Methods Mol. Biol.  119: 1
  6. Munari F. et al. (2012) J. Biol. Chem.  287: 33756
  7. Strelow J.M. et al. (2016) J. Biomol. Screen21: 786
  8. Wang H. et al. (2004) Nature 431: 873
 
Alternate Names
Nucleosome

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