Histone H2B, ubiquitylated (Lys120) (Histone H2B Type 1-B, Histone H2B.f, H2B/f, H2BFF, Histone H2B.1, H2B.1, Histone Cluster 1 H2bb, HIST1H2BB, MGC119804)

The nucleosome, made up of four core histone proteins (H2A, H2B, H3 and H4), is the primary\nbuilding block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation and ubiquitylation (1). Ubiquitin is a conserved 76 aa peptide unit that can be covalently\nlinked to many cellular proteins by the ubiquitylation process. Three components are involved in this protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thiolester complex with the activation component E1; the activated ubiquitin is subsequently transferred to the ubiquitin- carrier protein E2, then from E2 to ubiquitin ligase E3 for final delivery to the epsilon-NH2 of the target protein lysine residue (2). Histone H2B is mono-ubiquitylated on lysine 120 during transcriptional activation by the RAD6 E2 protein in conjunction with the BRE1A/BRE1B E3 ligase (also known as RNF20/RNF40) (3). The RAD6/BRE1 complex is recruited to gene promoters during activation by the PAF complex, an RNA polymerase II-associated protein complex that regulates transcriptional elongation (3-5). Mono-ubiquitylated\nhistone H2B lysine 120 is associated with the transcribed region of active genes (3,6). Mono-ubiquitylation of histone H2B stimulates transcriptional elongation by facilitating\nFACT-dependent chromatin remodeling (7,8). In addition, it is essential for subsequent methylation of histone H3 lysines 4 and 79, two additional histone modifications that regulate\ntranscriptional initiation and elongation (9). Interestingly, deubiquitylation of histone H2B lysine 120 by USP22, a subunit of the human SAGA histone acetyltransferase complex, is a required step in transcriptional activation (10). Thus, it appears that the ubiquitylation state of histone H2B is dynamic during transcription and may serve as an intermediate step in transcriptional activation.\n\nApplications: \nSuitable for use in Western Blot and Chromatin IP. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000 Incubate membrane with diluted antibody in 5% BSA, 1X TBS, 0.1% Tween-20 at 4 degrees C with gentle shaking, overnight.\nChromatin IP: 1:50\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

H5110-09U

Size

100ul

Applications

WB

Hosts

Rabbit

Reactivities

Hum, Mouse, Rat, NHP

Form

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, less than 0.02% sodium azide, 50% glycerol.

P Type

Mab

Purity

Purified

Isotype

IgG

References

(1) Peterson, C.L. and Laniel, M.A. (2004) Curr Biol 14, R546-51. (2) Liu, F. and Walters, K.J. (2010) Trends Biochem Sci 35,352-60. (3) Kim, J. et al. (2009) Cell 137, 459-71. (4) Wood, A. et al. (2003) J Biol Chem 278, 34739-42. (5) Xiao, T. et al. (2005) Mol Cell Biol 25, 637-51. (6) Minsky, N. et al. (2008) Nat Cell Biol 10, 483-8. (7) Pavri, R. et al. (2006) Cell 125, 703-17. (8) Fleming, A.B. et al. (2008) Mol Cell 31, 57-66. (9) Shilatifard, A. (2006) Annu Rev Biochem 75, 243-69. (10) Wyce, A. et al. (2004) Novartis Found Symp 259, 63-73; discussion 73-7, 163-9.

Additional Info

Recognizes endogenous levels of histone H2B, ubiquitylated on Lys120. Species Crossreactivity: monkey, mouse, rat.