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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
SQSTM1/p62 (D5E2) Rabbit mAb 8025 20 µl
Western Blotting Immunoprecipitation
H Mk 62 Rabbit IgG
TRAF6 (D21G3) Rabbit mAb 8028 20 µl
Western Blotting Immunoprecipitation
H Mk 60 Rabbit IgG
K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb 5621 20 µl
Western Blotting
All Rabbit IgG
TrkA (12G8) Rabbit mAb 2510 20 µl
Western Blotting Immunohistochemistry
H 140 Rabbit IgG
NRF2 (D1Z9C) XP® Rabbit mAb 12721 20 µl
Western Blotting Immunoprecipitation Flow Cytometry Chromatin Immunoprecipitation
H M Mk 97-100 Rabbit IgG
KEAP1 (D6B12) Rabbit mAb 8047 20 µl
Western Blotting Immunofluorescence
H M R 60-64 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
Western Blotting

Product 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.

Specificity / Sensitivity

Each antibody in the Sequestosome Signaling Antibody Sampler Kit detects endogenous levels of its target protein. K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb detects polyubiquitin chains formed by Lys63 residue linkage. It does not react with monoubiquitin or polyubiquitin chains formed by linkage to a different lysine residue. TRAF6 (D21G3) Rabbit mAb is not predicted to cross-react with other TRAF family members. TrkA (12G8) Rabbit mAb does not cross-react with TrkB.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly162 of human SQSTM1/p62 protein, residues near the amino terminus of human TRAF6 protein, residues surrounding the Lys63 branch of the human diubiquitin chain, residues surrounding Arg220 of human TrkA, residues surrounding Ala275 of human NRF2 protein, and residues near the carboxy terminus of human KEAP1 protein.

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).

1.  Geetha, T. et al. (2005) Mol Cell 20, 301-12.

2.  Kirkin, V. et al. (2009) Mol Cell 34, 259-69.

3.  Cullinan, S.B. et al. (2004) Mol Cell Biol 24, 8477-86.

4.  Seibenhener, M.L. et al. (2007) FEBS Lett 581, 175-9.

5.  Nguyen, T. et al. (2005) J Biol Chem 280, 32485-92.

6.  Segal, R.A. and Greenberg, M.E. (1996) Annu Rev Neurosci 19, 463-89.

7.  Komatsu, M. et al. (2010) Nat Cell Biol 12, 213-23.

8.  Jaiswal, A.K. (2004) Free Radic Biol Med 36, 1199-207.

9.  Bjørkøy, G. et al. Autophagy 2, 138-9.

10.  Joung, I. et al. (1996) Proc Natl Acad Sci U S A 93, 5991-5.

11.  Sanchez, P. et al. (1998) Mol Cell Biol 18, 3069-80.

12.  Puls, A. et al. (1997) Proc Natl Acad Sci U S A 94, 6191-6.

13.  Vadlamudi, R.K. et al. (1996) J Biol Chem 271, 20235-7.

14.  Wooten, M.W. et al. (2005) J Biol Chem 280, 35625-9.

15.  Wooten, M.W. et al. (2005) J Biol Chem 280, 35625-9.

16.  Bjørkøy, G. et al. (2005) J Cell Biol 171, 603-14.

17.  Komatsu, M. et al. (2007) Cell 131, 1149-63.

18.  Lee, D.F. et al. (2009) Mol Cell 36, 131-40.

19.  Pankiv, S. et al. (2007) J Biol Chem 282, 24131-45.

20.  Huang, E.J. and Reichardt, L.F. (2003) Annu Rev Biochem 72, 609-42.

Entrez-Gene Id 9817 , 4780 , 8878 , 7189 , 4914
Swiss-Prot Acc. Q14145 , Q16236 , Q13501 , Q9Y4K3 , P04629

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 5,675,063.

Sequestosome Signaling Antibody Sampler Kit