WB, IP, ChIP
M
Endogenous
75, 85
Rabbit IgG
#Q9EST8
80859
Product Information
Product Usage Information
For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:50 |
| Chromatin IP | 1:50 |
Storage
Specificity / Sensitivity
Species Reactivity:
Mouse
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly108 of mouse IκB-ζ protein.
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).
- Baeuerle, P.A. and Baltimore, D. (1988) Science 242, 540-6.
- Beg, A.A. and Baldwin, A.S. (1993) Genes Dev 7, 2064-70.
- Finco, T.S. et al. (1994) Proc Natl Acad Sci USA 91, 11884-8.
- Brown, K. et al. (1995) Science 267, 1485-8.
- Brockman, J.A. et al. (1995) Mol Cell Biol 15, 2809-18.
- Traenckner, E.B. et al. (1995) EMBO J 14, 2876-83.
- Chen, Z.J. et al. (1996) Cell 84, 853-62.
- Karin, M. and Ben-Neriah, Y. (2000) Annu Rev Immunol 18, 621-63.
- Yamazaki, S. et al. (2001) J Biol Chem 276, 27657-62.
- Kitamura, H. et al. (2000) FEBS Lett 485, 53-6.
- Haruta, H. et al. (2001) J Biol Chem 276, 12485-8.
- Matsuo, S. et al. (2007) Biochem J 405, 605-15.
- Totzke, G. et al. (2006) J Biol Chem 281, 12645-54.
- Motoyama, M. et al. (2005) J Biol Chem 280, 7444-51.
- Yamamoto, M. et al. (2004) Nature 430, 218-22.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
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