PathScan® Acetyl-Histone H4 Sandwich ELISA Kit #7238

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	Catalog #		Size & Concentration		Price(£)		Qty
	7238S		1 Kit (96 assays)		377.00
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Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer (10X), which is stored at –20°C (packaged separately).

Species Cross-Reactivity

H M Mk

Reactivity Key:  H=Human  M=Mouse  Mk=Monkey

Description

The PathScan® Acetyl-Histone H4 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of acetylated lysines on histone H4. A Histone H4 antibody has been coated onto the microwells. After incubation with cell lysates, Histone H4 is captured by the coated antibody. Following extensive washing, a Biotinylated Acetylated-Lysine Mouse mAb is added to detect the acetylated lysines on the Histone H4 protein. An HRP-linked Streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB is added to develop color. HRP substrate, TMB is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of acetylated Histone H4.

Specificity / Sensitivity

CST's PathScan® Acetyl-Histone H4 Sandwich ELISA Kit detects endogenous levels of acetylated histone H4. Using this Sandwich ELISA Kit #7238, acetylated lysines on Histone H4 are detected when treated with TSA in Jurkat cells. However, the levels of Histone H4 remain unchanged, as shown by Western analysis using the Histone H4 Antibody #2592 (figure 1). COS and NIH 3T3cells treated with TSA show similar results (data not shown).

ELISA - Western correlation

Figure 1: Treatment of Jurkat cells with TSA causes accumulation of acetylation on Histone H4, detected by Sandwich ELISA Kit #7238, but does not affect the level of total Histone H4 protein, detected by Western analysis. OD450nm readings are shown in the top figure, while the corresponding Western blot using the Acetylated Lysine mouse mAb (Ac-K-103) #9681 (left panel) or Histone H4 Antibody #2592 (right panel), is shown in the bottom figure.

Sensitivity

Figure 2: The relationship between protein concentration of lysates from untreated and TSA treated Jurkat cells and kit assay optical density readings. Jurkat cells were treated with TSA (0.4 µM overnight).

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, on 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 of 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 of H3 Thr3 in prophase and its dephosphorylation during anaphase (11).

1. Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79.
2. Hansen, J.C. et al. (1998) Biochemistry 37, 17637-41.
3. Strahl, B.D. and Allis, C.D. (2000) Nature 403, 41-5.
4. Cheung, P. et al. (2000) Cell 103, 263-71.
5. Bernstein, B.E. and Schreiber, S.L. (2002) Chem Biol 9, 1167-73.
6. Jaskelioff, M. and Peterson, C.L. (2003) Nat Cell Biol 5, 395-9.
7. Thorne, A.W. et al. (1990) Eur J Biochem 193, 701-13.
8. Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60.
9. Goto, H. et al. (1999) J Biol Chem 274, 25543-9.
10. Preuss, U. et al. (2003) Nucleic Acids Res 31, 878-85.
11. Dai, J. et al. (2005) Genes Dev 19, 472-88.

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7238:

Sandwich ELISA

• 2935 Histone H4 (L64C1) Mouse mAb
• 2592 Histone H4 Antibody
• 9681 Acetylated-Lysine Mouse mAb (Ac-K-103)
• 7233 PathScan® Acetyl-Histone H2A Sandwich ELISA Kit
• 7232 PathScan® Acetylated Histone H3 Sandwich ELISA Kit
• 7236 PathScan® Acetylated p53 Sandwich ELISA Kit
• 7178 PathScan® Acetyl-Histone H2B Sandwich ELISA Kit

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.