Figure 1. Treatment of COS cells with trichostatin A (TSA) increases the acetylation of Histone H3 at Lys18 detected by PathScan® Acetyl-Histone H3 (Lys18) Sandwich ELISA Kit #7122. TSA treatment does not affect the level of histone H3 that is detected by Western analysis. COS cells (70-80% confluent) were treated for 16-18 hours with 0.4 μM TSA at 37ºC. Absorbance readings at 450 nm are shown in the top figure while the corresponding Western blots using Histone H3 Antibody #9715 (left panel) or Acetyl-Histone H3 (Lys18) Antibody #9675 (right panel) are shown in the bottom figure.Learn more about how we get our images
Figure 2: The relationship between the protein concentration of the lysate from untreated and TSA-treated HeLa cells and kit assay optical density readings. HeLa cells were treated with TSA (4 μM overnight). An acid extraction was performed for cell lysis in the presence of 5mM sodium butyrate.Learn more about how we get our images
|Product Includes||Volume||Solution Color|
|Acetyl-Histone H3 (K18) Detection Ab||11 ml||Green|
|Anti-rabbit IgG, HRP-linked Antibody||11 ml||Red|
The PathScan® Acetyl-Histone H3 (Lys18) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of histone H3 when acetylated at Lys18. A Histone H3 Mouse Antibody* has been coated onto the microwells. After incubation with cell lysates, histone H3 protein is captured by the coated antibody. Following extensive washing, Acetyl-Histone H3 (Lys18) Rabbit Antibody* is added to detect histone H3 when acetylated at Lys18. 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 H3 acetylated at Lys18. *Antibodies in this kit are custom formulations specific to kit.
* Antibodies in kit are custom formulations specific to kit.
CST's PathScan® Acetyl-Histone H3 (Lys18) Sandwich ELISA Kit #7122 detects endogenous levels of histone H3 when acetylated at Lys18. As shown in Figure 1 using the Acetyl-Histone H3 (Lys18) Sandwich ELISA Kit #7122, a high level of acetylation at Lys18 on histone H3 is detected in COS cells when treated with TSA. The level of total histone H3 (modified and unmodified) remains unchanged as shown by Western analysis (Figure 1). Similar results are obtained when NIH/3T3 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.
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 post-translational 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).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. PathScan is a trademark of Cell Signaling Technology, Inc.
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