Apoptosis and Autophagy

SQSTM1/p62 Antibody #5114

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Applications Key:   W = Western Blotting  IF-IC = Immunofluorescence (Immunocytochemistry)
Reactivity Key: H = Human  M = Mouse  Mk = Monkey  R = Rat  
Species cross-reactivity is determined by Western blot.

Protocols

5114

Western Blotting, Immunofluorescence,

Specificity / Sensitivity

SQSTM1/p62 Antibody detects endogenous levels of total SQSTM1/p62 protein.

Source / Purification

Polyclonal antibodies were produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly410 of human SQSTM1/p62 protein. Antibodies were purified by protein A and peptide affinity chromatography.

IF-IC

Confocal immunofluorescent analysis of HeLa cells using SQSTM1/p62 Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Western Blotting

Western blot analysis of extracts from various cell lines using SQSTM1/p62 Antibody.

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with mouse SQSTM1 isoform c (mSQSTM1c) construct (+), using SQSTM1/p62 Antibody.

Background

Sequestosome 1 (SQSTM1, p62) is a ubiquitin binding protein that plays a role 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). It was subsequently found to interact with ubiquitin, providing a scaffold for several signaling proteins as well as 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 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 NFR2 activity.

1. Kirkin, V. et al. (2009) Mol Cell34, 259 - 69.
2. Seibenhener, M.L. et al. (2007) FEBS Lett581, 175 - 9.
3. Komatsu, M. et al. (2010) Nat Cell Biol12, 213 - 23.
4. Bjørkøy, G. et al. (2006) Autophagy2, 138 - 9.
5. Joung, I. et al. (1996) Proc Natl Acad Sci U S A93, 5991 - 5.
6. Sanchez, P. et al. (1998) Mol Cell Biol18, 3069 - 80.
7. Puls, A. et al. (1997) Proc Natl Acad Sci U S A94, 6191 - 6.
8. Vadlamudi, R.K. et al. (1996) J Biol Chem271, 20235 - 7.
9. Wooten, M.W. et al. (2005) J Biol Chem280, 35625 - 9.
10. Bjørkøy, G. et al. (2005) J Cell Biol171, 603 - 14.
11. Komatsu, M. et al. (2007) Cell131, 1149 - 63.
12. Pankiv, S. et al. (2007) J Biol Chem282, 24131 - 45.

Application References

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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.