Cell Cycle / Checkpoint Control
| CSTコード |
包装 |
希望納入価格（円） |
国内在庫  |
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| #2187S | 100 μL | 57,000 | |
|
CENPA抗体製品一覧
推奨プロトコール
最適な結果を得るために：Cell Signaling Technology (CST) 社は、各製品の推奨プロトコールを使用することを強くお薦めいたします。
推奨プロトコールはCST社内試験の徹底的なバリデーションに基づいて作成されておりますので、正確かつ再現性の高い結果が得られます。
注：各製品に最適化されたプロトコールをリンクしています。
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2187:
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Immunofluorescence
Immunoprecipitation
Western Blotting
| 用途（希釈倍率） | |
| ウエスタンブロッティング（1:1,000）、免疫沈降（1:25）、免疫蛍光細胞染色（IF-IC）（1:100） |
| 特異性・感度 | |
| 内在性レベルのSer7 がリン酸化されたヒトのCENP-A タンパク質を検出します。Histone H3 タンパク質などの他のHistone タンパク質とは交差しません。 |
| 使用抗原 | |
| ヒトのCENP-A タンパク質のSer7 周辺領域（合成リン酸化ペプチド） |
| ※括弧付きの動物種は配列が100%相同であるため反応すると推定されます。 |
Western Blotting
Western blot analysis of extracts from HeLa cells arrested in S phase or mitosis using Phospho-CENP-A (Ser7) Antibody (upper panel) or CENP-A Antibody #2186 (lower panel). S phase cells were treated for 12 hours with thymidine (2 mM), rinsed three times, released into normal growth medium for 10 hours and then treated an additional 12 hours with thymidine before harvesting. Mitotic cells were treated for 12 hours with thymidine, rinsed three times and then treated for 16 hours with paxitaxol (500 nM final).
IF-IC
Confocal immunofluorescent analysis of a mitotic HeLa cell using Phospho-CENP-A (Ser7) Antibody (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor
®
555 Conjugate) #2116 (red). Phospho-CENP-A signal is localized to bright spots in the metaphase plate. Blue pseudocolor = DRAQ5
®
#4084 (fluorescent DNA dye).
Modulation of chromatin structure plays a critical role in the regulation of transcription and replication of the eukaryotic genome. The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. In addition to the growing number of post-translational histone modifications regulating chromatin structure, cells can also exchange canonical histones with variant histones that can directly or indirectly modulate chromatin structure (1). CENP-A, also known as the chromatin-associated protein CSE4 (capping-enzyme suppressor 4-p), is an essential histone H3 variant that replaces canonical histone H3 in centromeric heterochromatin (2). The greatest divergence between CENP-A and canonical histone H3 occurs in the amino-terminal tail of the protein, which binds linker DNA between nucleosomes and facilitates proper folding of centromeric heterochromatin (3). The amino-terminal tail of CENP-A is also required for recruitment of other centromeric proteins (CENP-C, hSMC1, hZW10), proper kinetochore assembly and chromosome segregation during mitosis (4). Additional sequence divergence in the histone fold domain is responsible for correct targeting of CENP-A to the centromere (5). Many of the functions of CENP-A are regulated by phosphorylation (6,7). Aurora A-dependent phosphorylation of CENP-A on Ser7 during prophase is required for proper targeting of Aurora B to the inner centromere in prometaphase, proper kinetochore/microtubule attachment and proper alignment of chromosomes during mitosis (6).
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Jin, J. et al. (2005)
Trends Biochem Sci
30, 680-7.
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Ausió, J. (2006)
Brief Funct Genomic Proteomic
5, 228-43.
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Heit, R. et al. (2006)
Biochem Cell Biol
84, 605-18.
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Van Hooser, A.A. et al. (2001)
J Cell Sci
114, 3529-42.
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Black, B.E. et al. (2004)
Nature
430, 578-82.
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Kunitoku, N. et al. (2003)
Dev Cell
5, 853-64.
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Zeitlin, S.G. et al. (2001)
J Cell Biol
155, 1147-57.
