Cell Signaling Technology

Product Pathways - NF-kB Signaling

NF-κB1 p105/p50 (D7H5M) Rabbit mAb #12540

sc-1190   sc-7178  

No. Size Price
12540S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
12540 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse, Endogenous 50 Active form. 120 Precursor Rabbit IgG
IP 1:100
ChIP 1:50

Species cross-reactivity is determined by western blot.

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

Homology

Species predicted to react based on 100% sequence homology: Monkey,

Specificity / Sensitivity

NF-κB1 p105/p50 (D7H5M) Rabbit mAb recognizes endogenous levels of total NF-κB1 p105/p50 protein.

NF-κB1 p105/p50 (D7H5M) Rabbit mAb兔单抗能够检测内源NF-κB1 p105/p50总蛋白水平。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly415 of human NF-κB p105/p50 protein.

此单克隆抗体通过合成肽免疫动物制备,这种合成肽是人源NF-κB p105/p50蛋白Gly415周围的肽段。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (30 ng/ml) for 1 hour and either 10 μl of NF-κB1 p105/p50 (D7H5M) 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 IκBα Promoter Primers #5552, human IAP2 promoter primers, 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.染色质免疫沉淀法检测4 x 106 HeLa细胞中的交联染色质,细胞通过Human Tumor Necrosis Factor-α (hTNF-α) #8902 (30 ng/ml) 处理1小时或者使用 10 μl of NF-κB1 p105/p50 (D7H5M) Rabbit mAb 或 2 μl of Normal Rabbit IgG #2729处理。使用试剂盒是SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003。丰富的DNA通过使用SimpleChIP® Human IκBα Promoter Primers #5552, human IAP2 promoter primers, 和SimpleChIP® Human α Satellite Repeat Primers #4486引物进行Real-Time PCR来量化。每个样品中免疫沉淀的DNA的数量形成与总数量的相对值。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using NF-κB1 p105/p50 (D7H5M) Rabbit mAb. Western blot分析多种细胞系的细胞提取物,使用抗体是NF-κB1 p105/p50 (D7H5M) Rabbit mAb。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (10 ng/ml) for the indicated times, using NF-κB1 p105/p50 (D7H5M) Rabbit mAb.Western blot分析HeLa细胞的细胞提取物,未处理(-)或使用Human Tumor Necrosis Factor-α (hTNF-α) #8902 (10 ng/ml)按指示时间处理,使用抗体是NF-κB1 p105/p50 (D7H5M) Rabbit mAb。

Background

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 translocates 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)。

Following IKK-mediated phosphorylation of p105 NF-κB at multiple sites (Ser921, 923, 927, and 932) on its carboxy-terminus, SCF/β-TrCP mediated processing produces the 50 kDa active form p50 (12,13).

在IKK介导的p105 NF-κB1 C-末端多位点(Ser921, 923, 927和932)发生磷酸化后,SCFβ-TrCP介导产生50 kDa 蛋白的活性形式p50 (12,13)。

  1. Baeuerle, P.A. and Henkel, T. (1994) Annu Rev Immunol 12, 141-79.
  2. Baeuerle, P.A. and Baltimore, D. (1996) Cell 87, 13-20.
  3. Haskill, S. et al. (1991) Cell 65, 1281-9.
  4. Thompson, J.E. et al. (1995) Cell 80, 573-82.
  5. Whiteside, S.T. et al. (1997) EMBO J 16, 1413-26.
  6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
  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. Heissmeyer, V. et al. (2001) Mol Cell Biol 21, 1024-35.
  13. Orian, A. et al. (2000) EMBO J 19, 2580-91.

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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.

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