Revision 4

#7232Store at +4C

1 个试剂盒

(96 assays)

Species Cross Reactivity

H

UniProt ID:

#P68431

Entrez-Gene Id:

#8350

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
Histone H3 Mouse mAb Coated Microwells 18137 96 tests +4C
Acetylated-Lysine Rabbit Detection mAb 13968 1 ea Green (Lyophilized) +4C
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) 13272 1 ea Red (Lyophilized) +4C
Detection Antibody Diluent 13339 11 ml Green +4C
HRP Diluent 13515 11 ml Red +4C
TMB Substrate 7004 11 ml +4C
STOP Solution 7002 11 ml +4C
Sealing Tape 54503 2 ea +4C
ELISA Wash Buffer (20X) 9801 25 ml +4C
ELISA Sample Diluent 11083 25 ml Blue +4C
Cell Lysis Buffer (10X) 9803 15 ml -20C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

The PathScan® Acetylated Histone H3 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of acetylated lysines on Histone H3. A Histone H3 Antibody has been coated onto the microwells. After incubation with cell lysates, Histone H3 is captured by the coated antibody. Following extensive washing, an Acetylated-Lysine Rabbit mAb is added to detect the acetylated lysines on the Histone H3 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 optical density for this developed color is proportional to the quantity of acetylated Histone H3.

*Antibodies in kit are custom formulations specific to kit.

Specificity/Sensitivity

CST's PathScan® Acetylated Histone H3 Sandwich ELISA Kit detects endogenous levels of Acetylated Histone H3. Using this Sandwich ELISA Kit #7232, acetylated lysines on Histone H3 are detected when treated with TSA in Jurkat cells. However, the levels of Histone H3 remains unchanged, as shown by Western analysis using the Histone H3 Antibody #9715 (figure 1). COS and HIH 3T3 cells treated with TSA show similar results (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.

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).

  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.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PathScan is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    限制使用

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    Revision 4
    #7232

    PathScan® Acetylated Histone H3 Sandwich ELISA Kit

    PathScan® Acetylated Histone H3 Sandwich ELISA Kit: Image 1 Expand Image
    图 1:Jurkat 细胞用 TSA 处理会导致组蛋白 H3 乙酰化的聚集,使用 Sandwich ELISA kit #7232 可进行检测,但不影响蛋白质印迹分析检测的总组蛋白 H3 的水平。OD 450nm 读数如顶部图所示,使用 Acetylated Lysine Mouse mAb (Ac-K-103) #9681(左小图)或 Histone H3 Antibody #9715(右小图)检测到的相应蛋白质印迹如底部图所示。
    PathScan® Acetylated Histone H3 Sandwich ELISA Kit: Image 2 Expand Image
    图 2:未经处理和已经 TSA 处理的 Hela 细胞的裂解物蛋白浓度与使用试剂盒测得的光密度读数之间的关系。Hela 细胞用 TSA(4.0 µM,过夜)处理,裂解物用 Cell Lysis Buffer (10X) #9803 制备。