Cell Signaling Technology

Product Pathways - NF-kB Signaling

IκB-ζ (D4I7C) Rabbit mAb (ChIP Preferred) #76041

No. Size Price
76041S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
76041 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Mouse, Endogenous 75, 85 Rabbit IgG
IP 1:50
ChIP 1:50

Species cross-reactivity is determined by western blot.

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

Specificity / Sensitivity

IκB-ζ (D4I7C) Rabbit mAb (ChIP Preferred) recognizes endogenous levels of total mouse IκB-ζ protein. This antibody has weak reactivity for rat and human. Product #93726 is preferred for western blot.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly108 of mouse IκB-ζ protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from J774A.1 and Raw 264.7 cell lines, untreated (-) or treated with LPS (1 μg/ml, 4 hr; +) using IκB-ζ (D4I7C) Rabbit mAb (ChIP Preferred) (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length mouse IκB-ζ (mIκB-ζ; +) using IκB-ζ (D4I7C) Rabbit mAb (ChIP Preferred).

IP

IP

Immunoprecipitation of IκB-ζ from Raw 264.7 cells treated with LPS (1 μg/ml; 4 hr). Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is IκB-ζ (D4I7C) Rabbit mAb (ChIP Preferred). Western blot was performed using IκB-ζ (D4I7C) Rabbit mAb (ChIP Preferred). Blots were developed using a confirmation specific secondary antibody to avoid reactivity with IgG.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 J774A.1 cells treated with LPS #14011 (1μg/mL, 4hours) and either 10 µl of IκB-ζ (D4I7C) Rabbit mAb (Mouse Specific) 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® Mouse CXCL2 Promoter Primers #39609, mouse LCN2 promoter primers, and SimpleChIP® Mouse MYT-1 Promoter Primers #8985. 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.

Background

The NF-κB/Rel transcription factors are present in the cytosol in an inactive state complexed with the inhibitory IκB proteins (1-3). Activation occurs via phosphorylation of IκBα at Ser32 and Ser36 followed by proteasome-mediated degradation that results in the release and nuclear translocation of active NF-κB (3-7). IκBα phosphorylation and resulting Rel-dependent transcription are activated by a highly diverse group of extracellular signals including inflammatory cytokines, growth factors, and chemokines. Kinases that phosphorylate IκB at these activating sites have been identified (8).

IκB-ζ (MAIL, INAP) is a unique IκB family member homologous to Bcl-3 and induced by IL-1 and Toll-like receptor (TLR) ligands (9-11). Like other family members, it contains carboxyl terminal ankyrin-repeats responsible for interaction with NF-κB, particularly p50. Unlike classical IκB family members (α, β, ε) which inhibit NF-κB translocation and are rapidly degraded upon cytokine treatment, IκB-ζ is cytokine-inducible and localized to the nucleus where it regulates NF-κB DNA binding and transactivation (12-14). Induction of IκB-ζ is required for TLR/IL-1 induction of a subset of NF-κB target genes, including IL-6 (15). However, the IκB-ζ can also inhibit transactivation of other targets, such as TNF-α (14,15).

  1. Baeuerle, P.A. and Baltimore, D. (1988) Science 242, 540-6.
  2. Beg, A.A. and Baldwin, A.S. (1993) Genes Dev 7, 2064-70.
  3. Finco, T.S. et al. (1994) Proc Natl Acad Sci USA 91, 11884-8.
  4. Brown, K. et al. (1995) Science 267, 1485-8.
  5. Brockman, J.A. et al. (1995) Mol Cell Biol 15, 2809-18.
  6. Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
  7. Chen, Z.J. et al. (1996) Cell 84, 853-62.
  8. Karin, M. and Ben-Neriah, Y. (2000) Annu Rev Immunol 18, 621-63.
  9. Yamazaki, S. et al. (2001) J Biol Chem 276, 27657-62.
  10. Kitamura, H. et al. (2000) FEBS Lett 485, 53-6.
  11. Haruta, H. et al. (2001) J Biol Chem 276, 12485-8.
  12. Matsuo, S. et al. (2007) Biochem J 405, 605-15.
  13. Totzke, G. et al. (2006) J Biol Chem 281, 12645-54.
  14. Motoyama, M. et al. (2005) J Biol Chem 280, 7444-51.
  15. Yamamoto, M. et al. (2004) Nature 430, 218-22.

Application References

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