Revision 1
#26317
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W
Reactivity:
H M R
Sensitivity:
Endogenous
MW (kDa):
62
Source/Isotype:
Rabbit
UniProt ID:
#Q13501
Entrez-Gene Id:
8878
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Storage
Specificity/Sensitivity
Phospho-SQSTM1/p62 (Ser366) Antibody recognizes endogenous levels of SQSTM1/p62 protein only when phosphorylated at Ser366.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser366 of human SQSTM1/p62 protein. Antibodies are purified by peptide affinity chromatography.
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. 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.
TANK-binding kinase 1 (TBK1) is activated by von Hippel-Lindau (VHL) loss or hypoxia in cancer cells. TBK1 phosphorylates p62/SQSTM1 on Ser366, which is essential for p62 stability and kidney cancer cell proliferation (13).
Background References
- 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.
- Hu, L. et al. (2020) Cancer Discov 10, 460-475.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
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Revision 1
Western blot analysis of extracts from A-498 and 786-O cells, untreated (-) or treated with phosphatase (+), using Phospho-SQSTM1/p62 (Ser366) Antibody (upper), SQSTM1/p62 (D1Q5S) Rabbit mAb #39749 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).