Cell Signaling Technology

Product Pathways - NF-kB Signaling

IKK Isoform Antibody Sampler Kit #9966

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
IKKα Antibody #2682 40 µl W IP H M R Mk (B) 85 Rabbit
IKKβ Antibody #2684 40 µl W H Mk 87 Rabbit
IKKγ Antibody #2685 40 µl W IHC-P H M R Mk 48 Rabbit
IKKε Antibody #2690 40 µl W H M R 80 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

IKKα, IKKβ, IKKγ, and IKKε antibodies detect endogenous levels of total IKKα, IKKβ, IKKγ and IKKε proteins, respectively. These antibodies do not cross-react with IKK subunits other than their specified target.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3, HeLa and PC12 cells using IKKα Antibody #2682.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, NIH/3T3 and PC12 cells using IKKβ Antibody #2684.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using IKKγ Antibody #2685.


Western Blotting

Western Blotting

Western blot analysis of recombinant IKKε Kinase #7553 and Raw 264.7 cells, untreated or LPS-treated, using IKKε Antibody #2690.

Description

The IKK Isoform Antibody Sampler Kit provides an economical means to investigate NFkappaB signaling within the cell. The kit contains primary and secondary antibodies to perform four Western blots with each antibody.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues at the carboxyl terminus of human IKKβ, IKKε and IKKγ, to the 20 amino-terminal residues of human IKKα. Antibodies are purified by protein A and peptide affinity chromatography.

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 and IKKγ serves as the regulatory subunit (8,9). Activation of IKK depends upon phosphorylation of Ser177 and Ser181 in the activation loop of IKKβ (Ser176 and Ser180 in IKKα), which causes conformational changes resulting in kinase activation (10-13).

  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. 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

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

Companion Products

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

Products