Product Pathways - Autophagy Signaling
Phospho-SQSTM1/p62 (Thr269/Ser272) Antibody #13121
|13121S||100 µl (10 western blots)||---||In Stock||---|
|13121||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse, Rat||Endogenous||62||Rabbit|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation
Species predicted to react based on 100% sequence homology: Monkey.
Specificity / Sensitivity
Phospho-SQSTM1/p62 (Thr269/Ser272) Antibody recognizes endogenous levels of SQSTM1 protein only when phosphorylated at Thr269 and Ser272. This antibody may react with either dually or singly phosphorylated SQSTM1/p62. A background band is detected at 75 kDa in some cell lines.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr269/Ser272 of human SQSTM1/p62 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Western blot analysis of extracts from A549 cells, untreated (-) or treated with Roscovitine #9885 (20 μM, overnight; +), using Phospho-SQSTM1/p62 (Thr269/Ser272) Antibody (upper), SQSTM1/p62 (D5E2) Rabbit mAb #8025 (middle) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from chloroquine-treated HeLa cells (50 μM, overnight; +) and untreated (-) MCF7 cells, untreated (-) or treated with λ phosphatase and calf intestinal phosphatase (CIP) (+), using Phospho-SQSTM1/p62 (Thr269/Ser272) Antibody (upper) and SQSTM1/p62 (D5E2) Rabbit mAb #8025 (lower).
Immunoprecipitation of Phospho-SQSTM1/p62 (Thr269/Ser272) from ACHN cells using Normal Rabbit IgG #2729 (lane 2) or Phospho-SQSTM1/p62 (Thr262/Ser272) Antibody (lane 3). Lane 1 is 10% input. Western blot was performed using Phospho-SQSTM1/p62 (Thr262/Ser272) Antibody. Mouse Anti-rabbit IgG (Light-Chain Specific) (L57A3) mAb
#3677 was used as a secondary antibody to avoid cross-reaction with IgG heavy chain.
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. Studies have demonstrated a link between SQSTM1 and oxidative stress. SQSTM1 interacts with KEAP1, which is a cytoplasmic inhibitor of NRF2, a key transcription factor involved in cellular responses to oxidative stress (3). Thus, accumulation of SQSTM1 can lead to an increase in NRF2 activity.
Phosphorylation of SQSTM1 at Thr269 and Ser272 during mitosis by CDK1 can regulate cell cycle progression (13).
- 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. (2006) Autophagy 2, 138-9.
- Joung, I. et al. (1996) Proc Natl Acad Sci USA 93, 5991-5.
- Sanchez, P. et al. (1998) Mol Cell Biol 18, 3069-80.
- Puls, A. et al. (1997) Proc Natl Acad Sci USA 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.
- Linares, J.F. et al. (2011) Mol Cell Biol 31, 105-17.
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