CST, 86653S, Ubiquityl-Histone H2B (Lys120) (D11) Rabbit Monoclonal Antibody (PE Conjugate)

Monoclonal Antibody for studying H2B (Lys120) ubiquitinate. Validated for Flow Cytometry (Fixed/Permeabilized). Highly specific and rigorously validated in-house, Ubiquityl-Histone H2B (Lys120) (D11) Rabbit Monoclonal Antibody (PE Conjugate) (CST #86653) is ready to ship. Product Usage Information Flow Cytometry (Fixed/Permeabilized): 1:50 Storage Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze. Protocol Available protocols: Flow Cytometry (Fixed/Permeabilized) Specificity / Sensitivity Ubiquityl-Histone H2B (Lys120) (D11) Rabbit Monoclonal Antibody (PE Conjugate) detects endogenous levels of histone H2B protein only when ubiquitylated on Lys120. This antibody conjugate does not cross-react with other ubiquitylated proteins or free ubiquitin. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human histone H2B protein in which Lys120 is mono-ubiquitylated. Background The nucleosome, made up of four core histone proteins (H2A, H2B, H3 and H4), is the primary building 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 amino acid peptide unit that can be covalently linked 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-NH 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 histone H2B lysine 120 is associated with the transcribed region of active genes (3,6). Mono-ubiquitylation of histone H2B stimulates transcriptional elongation by facilitating FACT-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 transcriptional initiation and elongation (9). Interestingly, de-ubiquitylation 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. Alternate Names H2B; H2B clustered histone 3; H2B histone family, member F; H2B-clustered histone 3; H2B.1; H2B/f; H2B1B; H2BC3; H2BFF; HIST1H2BB; histone 1, H2bb; histone cluster 1 H2B family member b; histone cluster 1, H2bb; Histone H2B type 1-B; Histone H2B.1; Histone H2B.f; MGC119804 Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous Source/Isotype: Rabbit IgG