| Product Includes | Product # | Quantity | Mol. Wt | Isotype/Source |
|---|---|---|---|---|
| SQSTM1/p62 (D5E2) Rabbit mAb | 8025 | 20 µl | 62 kDa | Rabbit IgG |
| TRAF6 (D21G3) Rabbit mAb | 8028 | 20 µl | 60 kDa | Rabbit IgG |
| K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb | 5621 | 20 µl | Rabbit IgG | |
| TrkA (12G8) Rabbit mAb | 2510 | 20 µl | 140 kDa | Rabbit IgG |
| NRF2 (D1Z9C) XP® Rabbit mAb | 12721 | 20 µl | 97-100 kDa | Rabbit IgG |
| KEAP1 (D6B12) Rabbit mAb | 8047 | 20 µl | 60-64 kDa | Rabbit IgG |
| Anti-rabbit IgG, HRP-linked Antibody | 7074 | 100 µl | Goat |
Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.
Description
The Sequestosome Signaling Antibody Sampler Kit contains reagents to investigate sequestosome signaling within the cell. The kit contains enough antibodies to perform two western blot experiments per primary antibody.
Storage
Background
Sequestosome 1 (SQSTM1, p62) is a ubiquitin binding protein involved in cell signaling, oxidative stress, and autophagy (1-4). It was first identified as a protein that binds to the SH2 domain of p56Lck (5) and independently found to interact with PKCζ (6,7). SQSTM1 was subsequently found to interact with ubiquitin, providing a scaffold for several signaling proteins and triggering degradation of proteins through the proteasome or lysosome (8). Interaction between SQSTM1 and TRAF6 leads to the K63-linked polyubiquitination of TRAF6 and subsequent activation of the NF-κB pathway (9). Protein aggregates formed by SQSTM1 can be degraded by the autophagosome (4,10,11). SQSTM1 binds autophagosomal membrane protein LC3/Atg8, bringing SQSTM1-containing protein aggregates to the autophagosome (12). Lysosomal degradation of autophagosomes leads to a decrease in SQSTM1 levels during autophagy; conversely, autophagy inhibitors stabilize SQSTM1 levels. SQSTM1 also interacts with KEAP1, which is a cytoplasmic inhibitor of NRF2, a key transcription factor involved in cellular responses to oxidative stress (3). Under basal conditions, KEAP1 binds and retains NRF2 in the cytoplasm where it can be targeted for ubiquitin-mediated degradation (13). Small amounts of constitutive nuclear NRF2 maintain cellular homeostasis through regulation of basal expression of antioxidant response genes. Following oxidative or electrophilic stress, KEAP1 releases NRF2, thereby allowing the activator to translocate to the nucleus and bind to ARE-containing genes (14). The coordinated action of NRF2 and other transcription factors mediates the response to oxidative stress (15). Thus, accumulation of SQSTM1 can lead to an increase in NRF2 activity (3). KEAP1 also targets the down regulation of NF-κB activity by targeting IKKβ degradation (16). TrkA is a member of Trk receptor tyrosine kinases and is activated by NGF, which stimulates TrkA polyubiquitination (17,18). TrkA regulates proliferation and is important for development and maturation of the nervous system (19). SQSTM1 interaction with TRAF6 controls synthesis of K63 polyubiquititination on TrkA (18, 20). TrkA polyubiquitination is essential for neurotrophin-dependent receptor internalization, cell differentiation, and signaling (18).
- Kirkin, V. et al. (2009) Mol Cell 34, 259-69.
- Seibenhener, M.L. et al. (2007) FEBS Lett 581, 175-9.
- Komatsu, M. et al. (2010) Nat Cell Biol 12, 213-23.
- Bjørkøy, G. et al. Autophagy 2, 138-9.
- Joung, I. et al. (1996) Proc Natl Acad Sci U S A 93, 5991-5.
- Sanchez, P. et al. (1998) Mol Cell Biol 18, 3069-80.
- Puls, A. et al. (1997) Proc Natl Acad Sci U S A 94, 6191-6.
- Vadlamudi, R.K. et al. (1996) J Biol Chem 271, 20235-7.
- Wooten, M.W. et al. (2005) J Biol Chem 280, 35625-9.
- Bjørkøy, G. et al. (2005) J Cell Biol 171, 603-14.
- Komatsu, M. et al. (2007) Cell 131, 1149-63.
- Pankiv, S. et al. (2007) J Biol Chem 282, 24131-45.
- Cullinan, S.B. et al. (2004) Mol Cell Biol 24, 8477-86.
- Nguyen, T. et al. (2005) J Biol Chem 280, 32485-92.
- Jaiswal, A.K. (2004) Free Radic Biol Med 36, 1199-207.
- Lee, D.F. et al. (2009) Mol Cell 36, 131-40.
- Huang, E.J. and Reichardt, L.F. (2003) Annu Rev Biochem 72, 609-42.
- Geetha, T. et al. (2005) Mol Cell 20, 301-12.
- Segal, R.A. and Greenberg, M.E. (1996) Annu Rev Neurosci 19, 463-89.
- Wooten, M.W. et al. (2005) J Biol Chem 280, 35625-9.
Background References
Trademarks and Patents
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
