Phospho-TAB2 (Ser372) (D5A4) R

8155:

Immunoprecipitation , Western Blotting

Phospho-TAB2 (Ser372) (D5A4) Rabbit mAb recognizes endogenous levels of TAB2 protein only when phosphorylated at Ser372.

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser372 of human TAB2 protein.

Western Blotting

Western blot analysis of extracts from 293T cells, untreated or treated with Human Interleukin-1β (hIL-1β) #8900 (50 ng/ml) and Calyculin A (Serine/Threonine Phosphatase Inhibitor) #9902 (CalA) (100 nM, 20 min), using Phospho-TAB2 (Ser372) (D5A4) Rabbit mAb (upper) or total TAB2 (C88H10) Rabbit mAb #3745 (lower). Lysates were then treated with a combination of calf intestinal phosphatase (CIP) and λ-phosphatase to show phospho-specificity.

TAK1 is a mitogen-activated protein kinase kinase kinase activated by TGF-β and various pro-inflammatory signals (1,2). In vivo , TAK1 activation requires its association with TAK1 binding protein 1 (TAB1), which triggers TAK1 autophosphorylation at Thr184 and Thr187 (3,4). The TAB2 adaptor protein links TAK1 with TRAF6 to mediate TAK1 activation following IL-1 stimulation (5). Once activated, TAK1 phosphorylates the MAPK kinases MKK4 and MKK3/6, which activate JNK and p38 MAPK, respectively. TAK1 and TRAF6 also activate the NF-κB pathway by phosphorylating the NF-κB inducing kinase (NIK) to trigger subsequent activation of IKK (2,6). In addition to TAK1, TAB1 interacts with and activates p38α MAPK (7). Targeted disruption of the TAB1 gene in mice causes a drastic reduction in TAK1 activity and leads to embryonic lethality (8).

TAK1 is associated with TAB1 as well as either TAB2 or TAB3 (9). Activation of TAK1 is triggered by K63-ubiquitination of TRAF6, resulting in binding to TAB2 and TAB3 and autophosphorylation of TAK1 (10-12). Multiple phosphorylation sites have been identified in all three TAB family members that are triggered by IL-1 (13,14). TAB2 phosphorylation was identified at Ser372 and Ser524 (14).

1. Yamaguchi, K. et al. (1995) Science 270, 2008-2011.
2. Ninomiya-Tsuji, J. et al. (1999) Nature 398, 252-256.
3. Shibuya, H. et al. (1996) Science 272, 1179-1182.
4. Sakurai, H. et al. (2000) FEBS Lett. 474, 141-145.
5. Takaesu, G. et al. (2000) Mol. Cell 4, 649-658.
6. Wang, C. et al. (2001) Nature 412, 346-351.
7. Ge, B. et al. (2002) Science 295, 1291-1294.
8. Komatsu, Y. et al. (2002) Mech. Dev. 119, 239-249.
9. Cheung, P.C. et al. (2004) Biochem J 378, 27-34.
10. Kishida, S. et al. (2005) Genes Cells 10, 447-54.
11. Sato, Y. et al. (2009) EMBO J 28, 3903-9.
12. Kanayama, A. et al. (2004) Mol Cell 15, 535-48.
13. Cheung, P.C. et al. (2003) EMBO J 22, 5793-805.
14. Mendoza, H. et al. (2008) Biochem J 409, 711-22.

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• 3745 TAB2 (C88H10) Rabbit mAb
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

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