PathScan® Acetyl-Histone H2B (Lys5) Sandwich ELISA Kit

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ELISA

Inquiry Info. # 7218

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Supporting Data

REACTIVITY

H M Mk

Application Key:

ELISA-ELISA

Species Cross-Reactivity Key:

H-Human
M-Mouse
Mk-Monkey

Product Information

Product Description

The PathScan^® Acetyl-Histone H2B (Lys5) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of histone H2B when acetylated at Lys5. A Histone H2B Antibody has been coated onto the microwells. After incubation with cell lysates, histone H2B protein (acetylated and non-acetylated) is captured by the coated antibody. Following extensive washing, Acetyl-Histone H2B (Lys5) Antibody is added to detect acetylated Lys5 on the histone H2B protein. Anti-rabbit IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of histone H2B acetylated at Lys5.
Antibodies in kit are custom formulations specific to kit.

Protocol

Specificity / Sensitivity

CST's PathScan^® Acetyl-Histone H2B (Lys5) Sandwich ELISA Kit detects endogenous levels of histone H2B when acetylated at Lys5. As shown in Figure 1 using the Acetyl-Histone H2B (Lys5) Sandwich ELISA Kit #7218, a high level of acetylation at Lys5 on histone H2B is detected in NIH/3T3 cells when treated with trichostatin A (TSA). The level of total histone H2B (acetylated and non-acetylated) remains unchanged as shown by Western analysis (Figure 1). Similar results are obtained when COS and Jurkat cells are treated with TSA (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Species Reactivity:

Human, Mouse, Monkey

Background

Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various posttranslational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11).

Database Links

UniProt ID: P33778

Entrez-Gene Id: 3018

Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.

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PathScan® Acetyl-Histone H2B (Lys5) Sandwich ELISA Kit #7218