Cell Signaling Technology

Product Pathways - Apoptosis

SQSTM1/p62 (D10E10) Rabbit mAb (IF Preferred) #7695

Applications Reactivity Sensitivity MW (kDa) Isotype
IP IF-IC H Endogenous 62 Rabbit IgG

Applications Key:  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

SQSTM1/p62 (D10E10) Rabbit mAb (IF Preferred) is recommended to detect endogenous levels of total SQSTM1/p62 protein by immunofluorescence. Products SQSTM1/p62 (D5E2) Rabbit mAb #8025 and SQSTM1/p62 Antibody #5114 are preferred for western blot.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human SQSTM1/p62 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using SQSTM1/p62 (D10E10) Rabbit mAb (IF Preferred).

Western Blotting

Western Blotting

Western blot analysis of extracts from SK-MEL-2 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® SQSTM1/p62 siRNA II #6399 (+), using SQSTM1/p62 (D10E10) Rabbit mAb (IF Preferred) (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The SQSTM1/p62 (D10E10) Rabbit mAb (IF Preferred) confirms silencing of SQSTM1/p62 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.

Western Blotting

Western Blotting

Western blot analysis of extracts from SK-MEL-2 cells, untreated (-) or starved overnight in Earle's Balanced Salt Solution (EBSS) (+), using SQSTM1/p62 (D10E10) Rabbit mAb (IF Preferred).


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with bafilomycin A (100 nM, 18 hr; right), using SQSTM1/p62 (D10E10) Rabbit mAb (IF Preferred) (green). Actin filaments were labeled with DY-554 phalloidin. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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.

  1. Kirkin, V. et al. (2009) Mol Cell 34, 259-69.
  2. Seibenhener, M.L. et al. (2007) FEBS Lett 581, 175-9.
  3. Komatsu, M. et al. (2010) Nat Cell Biol 12, 213-23.
  4. Bjørkøy, G. et al. (2006) Autophagy 2, 138-9.
  5. Joung, I. et al. (1996) Proc Natl Acad Sci USA 93, 5991-5.
  6. Sanchez, P. et al. (1998) Mol Cell Biol 18, 3069-80.
  7. Puls, A. et al. (1997) Proc Natl Acad Sci USA 94, 6191-6.
  8. Vadlamudi, R.K. et al. (1996) J Biol Chem 271, 20235-7.
  9. Wooten, M.W. et al. (2005) J Biol Chem 280, 35625-9.
  10. Bjørkøy, G. et al. (2005) J Cell Biol 171, 603-14.
  11. Komatsu, M. et al. (2007) Cell 131, 1149-63.
  12. Pankiv, S. et al. (2007) J Biol Chem 282, 24131-45.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

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

Products