Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-Tpl2 (Ser400) Antibody #4491

Applications Reactivity Sensitivity MW (kDa) Source
W R (H) (M) Transfected Only 60, 62 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-Tpl2 (Ser400) Antibody detects transfected Tpl2 only when phosphorylated at Ser400. This antibody does not cross-react with other phosphorylated MAP kinase kinase kinases.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a mixture synthetic phosphorylated peptides corresponding to residues surrounding Ser400 of human, mouse and rat Tpl2. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untransfected or transfected with rat Tpl2 , using Phospho-Tpl2 (Ser400) Antibody (upper) and Tpl2 Antibody #4492 (lower). In lane 3, Tpl2 transfected COS cell lysate was treated with λ phosphatase to demonstrate the phospho-specificity of Phospho-Tpl2 (Ser400) Antibody.

Background

Tpl2 (tumor progression locus 2), also known as COT (cancer osaka thyroid), is a serine/threonine kinase expressed primarily in hematopoietic tissues, lung and liver (1). Over-expression of Tpl2 potentiates MAP kinase pathways through MEK1 and SEK1, as well as through MKK6 and MEK5 (2,3). Tpl2 is also engaged in NF-κB activation through NF-κB inducing kinase (NIK), or by inducing phosphorylation and degradation of the NF-κB precusor, p105 NF-κB1 (4,5). Ser400 of Tpl2 is phosphorylated in an Akt-dependent manner. This phosphorylation is required for Tpl2-induced NF-κB-dependent transcription (6). Tpl2 also activates caspase-3 by promoting the assembly of a protein complex of Apaf1 (apoptotic protease-activating factor 1), caspase-9, Tpl2, adaptor protein Tvl1 and procaspase-3 (7).

  1. Patriotis, C. et al. (1993) Proc Natl Acad Sci U S A 90, 2251-5.
  2. Salmeron, A. et al. (1996) EMBO J 15, 817-26.
  3. Chiariello, M. et al. (2000) Mol Cell Biol 20, 1747-58.
  4. Lin, X. et al. (1999) Immunity 10, 271-80.
  5. Belich, M.P. et al. (1999) Nature 397, 363-8.
  6. Kane, L.P. et al. (2002) Mol Cell Biol 22, 5962-74.
  7. Patriotis, C. et al. (2001) J Cell Physiol 187, 176-87.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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