Cell Signaling Technology

Product Pathways - NF-kB Signaling

Phospho-NF-κB p65 (Ser468) Antibody #3039

nf-kappab   NF-kB   NFkappaB   NFkB   nfκb p65   nuclear factor kappa B   Rel   RelA  

No. Size Price
3039S 100 µl ( 10 western blots ) ¥4,050.00 现货查询 购买询价 防伪查询
3039 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat, Endogenous 65 Rabbit
IP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation,

Specificity / Sensitivity

Phospho-NF-kappaB p65 (Ser468) Antibody detects NF-kappaB p65 only when phosphorylated at serine 468.Phospho-NF-kappaB p65 (Ser468) Antibody只能检测内源的在ser468位点磷酸化的NF-kappaB p65蛋白。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser468 of human NF-kappaB p65. Antibodies are purified by protein A and peptide affinity chromatography.

此多克隆抗体是通过合成人源对应的NF-kappaB p65 Ser468位点周围的磷肽段来免疫动物而获得。抗体是通过protein A和多肽亲和层析法纯化。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells treated for 5 minutes with TNF-alpha #2169 (20 ng/ml), Calyculin A #9902 (50 nM), or both compounds, using Phospho-NF-kappaB p65 (Ser468) Antibody (top) or NF-kappaB p65 Antibody #3034 (bottom).Western免疫印迹。用Phospho-NF-kappaB p65 (Ser468) Antibody (上图) 或 NF-kappaB p65 Antibody #3034 (下图)研究经TNF-alpha #2169 (20 ng/ml,5min)或Calyculin A #9902 (50 nM)或者经过两种试剂处理的HeLa细胞的细胞提取液。


Transcription factors of the nuclear factor κ B (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50), and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which is then translocated to the nucleus (9-11).

核因子κ B(NF-κB)/Rel 家族的转录调控因子在炎症反应和免疫反应中发挥了至关重要的作用(1,2)。在哺乳动物中一共有5个家族成员: RelA, c-Rel, RelB, NF-κB1 (p105/p50) 和 NF-κB2 (p100/p52)。 p105 和 p100 在蛋白水解酶的作用下分别形成p50 和 p52。Rel与p50和p52形成二聚体,此复合体能够结合到DNA上调控转录。在未刺激的状态下, NF-κB 在IκB抑制剂作用下在细胞质中处于非活性状态(3-5)。 NF-κB激活因子能够诱导 IκB 蛋白的磷酸化, 这就使IκB能够快速的经过泛素化-蛋白酶通路降解从而释放 NF-κB,激活的NF-κB入核调控基因的表达(6-8)。 NIK 和 IKKα (IKK1) 调节磷酸化并促使NF-κB2 (p100) 生成p52, p52之后会入核发挥功能(9-11)。

PMA-induced NF-kappaB transcriptional activity is dependent on the region between amino acids 442 and 470, suggesting a role for one or more of the potential phosphorylation sites (Ser457, Thr458, Thr464, or Ser468) in this region (12). T-cell costimulation and Calyculin A have both been shown to increase Ser468 phosphorylation (13, 14). IKKβ (but not IKKα) and GSK-3β both target this site (14, 15), which appears to have a negative regulatory role not involving inhibition of nuclear translocation after TNFα or IL-1β stimulation (15).

PMA诱导的NF-kappaB 转录活性是依靠442和470位点之间的区域,这就提示了这个区域几个潜在的磷酸化位点(Ser457, Thr458, Thr464和 Ser468)的功能(12)。T细胞刺激和Calyculin A能增加Ser468位点的磷酸化(13, 14)。IKKβ (不是IKKα) 和 GSK-3β的靶标都是这个位点 (14, 15), 他们具有负调控的作用,但是不参与到TNFα或 IL-1β刺激的核转录(15)。

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  4. Thompson, J.E. et al. (1995) Cell 80, 573-82.
  5. Whiteside, S.T. et al. (1997) EMBO J 16, 1413-26.
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  7. Scherer, D.C. et al. (1995) Proc Natl Acad Sci USA 92, 11259-63.
  8. Chen, Z.J. et al. (1996) Cell 84, 853-62.
  9. Senftleben, U. et al. (2001) Science 293, 1495-9.
  10. Coope, H.J. et al. (2002) EMBO J 21, 5375-85.
  11. Xiao, G. et al. (2001) Mol Cell 7, 401-9.
  12. Schmitz, M.L. et al. (1995) J Biol Chem 270, 15576-84.
  13. Mattioli, I. et al. (2004) Blood 104, 3302-4.
  14. Buss, H. et al. (2004) J Biol Chem 279, 49571-4.
  15. Schwabe, R.F. and Sakurai, H. (2005) FASEB J 19, 1758-60.

Application References

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