Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

SPT16 (D7I2K) Rabbit mAb #12191

FACT   FACT140   SUPT16H  

No. Size Price
12191S 100 µl ( 10 western blots ) ¥3,250.00 现货查询 购买询价 防伪查询
12191 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 140 Rabbit IgG
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, ChIP=Chromatin IP,


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

Specificity / Sensitivity

SPT16 (D7I2K) Rabbit mAb recognizes endogenous levels of total SPT16 protein.

SPT16 (D7I2K) Rabbit mAb 兔单抗能够检测内源性SPT16总蛋白水平。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu662 of human SPT16 protein.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using SPT16 (D7I2K) Rabbit mAb.Western blot 方法检测不同细胞系的提取物,使用的抗体为SPT16 (D7I2K) Rabbit mAb。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HCT 116 cells starved for 48 hr then serum stimulated with 10% FBS for 15 min and either 10 μl of SPT16 (D7I2K) 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 c-Fos Promoter Primers #4663, SimpleChIP® Human c-Fos Exon 3 Primers #12010, SimpleChIP® Human EGR1 Promoter Primers #5549, and SimpleChIP® Human EGR1 Intron 3 Primers #11953. 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 HCT 116细胞中提取的交联过的染色质,以及10 µl SPT16 (D7I2K) Rabbit mAb 或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀,这些细胞需要饥饿培养48小时然后用10% FBS进行15分钟血清刺激。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 c-Fos Promoter Primers #4663, SimpleChIP® Human c-Fos Exon 3 Primers #12010, SimpleChIP® Human EGR1 Promoter Primers #5549, 以及SimpleChIP® Human EGR1 Intron 3 Primers #11953。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。


Supressor of Ty-16 (SPT16) and structure-specific recognition protein-1 (SSRP1) are subunits of the facilitates chromatin transcription (FACT) complex that is essential for transcription elongation (1,2). FACT facilitates RNA polymerase-dependent transcription of chromatin templates by destabilizing the nucleosomes within the open reading frames of active genes (3-5). FACT destabilizes the nucleosomes, which would otherwise act as barriers to RNA polymerase transcription activity, by disrupting histone-histone and histone-DNA contacts that lead to the eviction of the histone H2A-H2B dimer (2,3,6). FACT may also function as a histone chaperone to reassemble nucleosomes after RNA polymerase passage (7). In addition to transcription, FACT activity has been shown to have a role in DNA replication in yeast and in DNA repair by contributing to the activation of p53 by CK2 and by facilitating histone H2AX-H2B exchange upon DNA damage (8-10).

Supressor of Ty-16 (SPT16)以及structure-specific recognition protein-1 (SSRP1)是转录延伸中至关重要的染色质转录促进复合物的亚单位(1,2)。FACT能够通过对活跃基因开放阅读框内核小体的去稳定作用来促进RNA聚合酶依赖的转录(3-5)。FACT能够破坏导致H2A-H2B二聚体丢失的组蛋白-组蛋白以及组蛋白-DNA的联系,从而使成为RNA聚合酶转录激活障碍的核小体去稳定(2,3,6)。FACT可能还可作为组蛋白伴侣在RNA聚合酶通路之后重新组装核小体(7)。除了转录,FACT还可以通过CK2促进p53活化以及促进DNA损伤的组蛋白H2AX-H2B交换,从而在酵母的DNA复制和DNA修复中发挥作用(8-10)。

  1. Winkler, D.D. and Luger, K. (2011) J Biol Chem 286, 18369-74.
  2. Orphanides, G. et al. (1999) Nature 400, 284-8.
  3. Orphanides, G. et al. (1998) Cell 92, 105-16.
  4. Birch, J.L. et al. (2009) EMBO J 28, 854-65.
  5. Orphanides, G. and Reinberg, D. (2000) Nature 407, 471-5.
  6. Keller, D.M. and Lu, H. (2002) J Biol Chem 277, 50206-13.
  7. Belotserkovskaya, R. et al. (2003) Science 301, 1090-3.
  8. Schlesinger, M.B. and Formosa, T. (2000) Genetics 155, 1593-606.
  9. Heo, K. et al. (2008) Mol Cell 30, 86-97.

Application References

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