Product Pathways - NF-kappaB Signaling

Phospho-IKK-alpha/beta (Ser176/180) Blocking Peptide #1023

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Description

This peptide is used to block Phospho-IKK-a/b (Ser176/180) (16A6) Rabbit mAb #2697 reactivity.

Quality Control

The quality of the peptide was evaluated by reverse-phase HPLC and by mass spectrometry. The peptide blocks Phospho-IKK-alpha/beta (Ser176/180) (16A6) Rabbit mAb #2697 signal in immunohistochemistry.

Applications

Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry protocols.

Directions for Use

For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 ul total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the relevant product data sheet.

Background

The NF-κB/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IκB proteins (1-3). Most agents that activate NF-κB do so through a common pathway based on phosphorylation-induced, proteasome-mediated degradation of IκB (3-7). The key regulatory step in this pathway involves activation of a high molecular weight IκB kinase (IKK) complex, whose catalysis is generally carried out by three tightly associated IKK subunits. IKKα and IKKβ serve as the catalytic subunits of the kinase. IKKγ serves as the regulatory subunit (8,9). Activation of IKK depends on phosphorylation; Ser177 and Ser181 in the activation loop of IKKβ (176 and 180 in IKKα) are the specific sites whose phosphorylation causes conformational changes resulting in kinase activation (10-13).

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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. Zandi, E. et al. (1997) Cell 91, 243-52.
9. Karin, M. (1999) Oncogene 18, 6867-74.
10. DiDonato, J.A. et al. (1997) Nature 388, 548-54.
11. Mercurio, F. et al. (1997) Science 278, 860-6.
12. Johnson, L.N. et al. (1996) Cell 85, 149-58.
13. Delhase, M. et al. (1999) Science 284, 309-13.

Application References

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Companion Products

• 2697 Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.