Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Histone H2A (D6O3A) Rabbit mAb #12349

No. Size Price
12349S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
12349 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey,Zebrafish, Endogenous 14 Rabbit IgG
IF-IC 1:200
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,


Species predicted to react based on 100% sequence homology: Hamster, Bovine, Dog, Guinea Pig,

Specificity / Sensitivity

Histone H2A (D6O3A) Rabbit mAb recognizes endogenous levels of total histone H2A protein. This antibody does not cross-react with other histone proteins.

Histone H2A (D6O3A) Rabbit mAb兔单抗能够检测内源性组蛋白H2A总蛋白水平。该抗体与其他组蛋白没有交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the carboxy terminus of human histone H2A protein.




Confocal immunofluorescent analysis of HeLa cells using Histone H2A (D6O3A) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).采用共聚焦免疫荧光术检测Hela细胞,使用的抗体为Histone H2A (D6O3A) Rabbit mAb (绿色)。肌动蛋白微丝使用DY-554 phalloidin进行标记(红色)。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H2A (D6O3A) Rabbit mAb.Western blot方法检测多种细胞系的提取物,使用的抗体为Histone H2A (D6O3A) Rabbit mAb。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 5 μl of Histone H2A (D6O3A) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MYT-1 Exon 1 Primers #4493, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.使用从4 x 106 MCF7细胞中提取的交联过的染色质,以及5 µl Histone H2A (D6O3A) Rabbit mAb 或2 µl Normal Rabbit IgG #2729进行染色质免疫沉。所用试剂盒为SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003。富集到的DNA经过实时PCR定量,所使用引物为SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human MYT-1 Exon 1 Primers #4493, 以及SimpleChIP® Human α Satellite Repeat Primers #4486。每个样品中免疫沉淀得到的DNA量由与input chromatin( 相当于1)的相对信号值来表示。


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).

染色质结构调整在真核生物的转录调控中扮演着一个重要角色。核小体是染色质的基本结构单位,由DNA缠绕八个核心组蛋白 (H2A、H2B、H3、H4各2个分子)组成(1)。核心组蛋白的氨基端尾部常发生多种翻译后修饰,包括乙酰化、磷酸化、甲基化和泛素化 (2-5)。这些修饰都是针对不同刺激而产生的反应,且会直接影响染色质对转录因子的可接近性,进而影响基因表达(6)。在多数物种中,组蛋白H2B 乙酰化主要发生在Lys5, 12, 15和20位点上 (4,7)。而组蛋白H3的乙酰化则主要发生在Lys9, 14, 18, 23, 27和56位点上。在一些生物体内,H3 Lys9位点的乙酰化修饰看似对组蛋白沉积和染色质组装有决定性的作用 (2,3)。组蛋白H3 Ser10、Ser28和Thr11位点上的磷酸化修饰与有丝分裂和减数分裂期间的染色体浓缩紧密相关(8-10)。组蛋白H3 Thr3位点上的磷酸化在许多物种中是高度保守的,由激酶haspin催化完成。在哺乳动物细胞内用磷酸化特异性抗体进行免疫染色,发现有丝分裂中H3 Thr3 位点的磷酸化发生在前期,而去磷酸化则发生在后期(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.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

用户评论 --- 共 0


我要参与评论 :