Anti-SQSTM1 / p62 antibody (ab155686)

Rabbit polyclonal SQSTM1 / p62 antibody. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human, Zebrafish. Cited in 7 publication(s). Independently reviewed in 3 review(s).

Overview

  • Product name

    Anti-SQSTM1 / p62 antibody
    See all SQSTM1 / p62 primary antibodies
  • Description

    Rabbit polyclonal to SQSTM1 / p62
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, Flow Cyt, IP, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Zebrafish
  • Immunogen

    Synthetic peptide corresponding to Human SQSTM1/ p62 aa 157-251 (internal sequence). Carrier-protein conjugated synthetic peptide.
    Database link: Q13501

  • Positive control

    • ICC/IF: HeLa cells treated with 50µM Chloroquine for 24 hr, HepG2 cells treated with 3µM thapsigargin 12 hrs. WB: HepG2 whole cell extracts, A549 whole cell lysate, HepG2 treated with 3µM Thapsigargin for 24 hours whole cell extracts, PC-12 whole cell lysate/extract, Rat2 whole cell lysate/extract. IP: HeLa whole cell extracts. IHC-P: human ovarian carcinoma tissue

Properties

Applications

Our Abpromise guarantee covers the use of ab155686 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/100 - 1/1000.
Flow Cyt 1/50 - 1/200.
IP 1/100 - 1/500.
WB 1/500 - 1/3000. Predicted molecular weight: 47 kDa.
ICC/IF 1/100 - 1/1000.

Target

  • Function

    Adapter protein which binds ubiquitin and may regulate the activation of NFKB1 by TNF-alpha, nerve growth factor (NGF) and interleukin-1. May play a role in titin/TTN downstream signaling in muscle cells. May regulate signaling cascades through ubiquitination. Adapter that mediates the interaction between TRAF6 and CYLD (By similarity). May be involved in cell differentiation, apoptosis, immune response and regulation of K(+) channels.
  • Tissue specificity

    Ubiquitously expressed.
  • Involvement in disease

    Defects in SQSTM1 are a cause of Paget disease of bone (PDB) [MIM:602080]. PDB is a metabolic bone disease affecting the axial skeleton and characterized by focal areas of increased and disorganized bone turn-over due to activated osteoclasts. Manifestations of the disease include bone pain, deformity, pathological fractures, deafness, neurological complications and increased risk of osteosarcoma. PDB is a chronic disease affecting 2 to 3% of the population above the age of 40 years.
  • Sequence similarities

    Contains 1 OPR domain.
    Contains 1 UBA domain.
    Contains 1 ZZ-type zinc finger.
  • Domain

    The UBA domain binds specifically 'Lys-63'-linked polyubiquitin chains of polyubiquitinated substrates. Mediates the interaction with TRIM55.
    The OPR domain mediates homooligomerization and interactions with PRKCZ, PRKCI, MAP2K5 and NBR1.
    The ZZ-type zinc finger mediates the interaction with RIPK1.
  • Post-translational
    modifications

    Phosphorylated. May be phosphorylated by PRKCZ (By similarity). Phosphorylated in vitro by TTN.
  • Cellular localization

    Cytoplasm. Late endosome. Nucleus. Sarcomere (By similarity). In cardiac muscles localizes to the sarcomeric band (By similarity). Localizes to late endosomes. May also localize to the nucleus. Accumulates in neurofibrillary tangles and in Lewy bodies of neurons from individuals with Alzheimer and Parkinson disease respectively. Enriched in Rosenthal fibers of pilocytic astrocytoma. In liver cells, accumulates in Mallory bodies associated with alcoholic hepatitis, Wilson disease, indian childhood cirrhosis and in hyaline bodies associated with hepatocellular carcinoma.
  • Information by UniProt
  • Database links

  • Alternative names

    • A170 antibody
    • DMRV antibody
    • EBI 3 associated protein of 60 kDa antibody
    • EBI 3 associated protein p60 antibody
    • EBI3 associated protein of 60 kDa antibody
    • EBI3 associated protein p60 antibody
    • EBI3-associated protein of 60 kDa antibody
    • EBIAP antibody
    • FTDALS3 antibody
    • MGC127197 antibody
    • ORCA antibody
    • OSF-6 antibody
    • Osi antibody
    • OSIL antibody
    • Oxidative stress induced like antibody
    • p60 antibody
    • p62 antibody
    • p62B antibody
    • Paget disease of bone 3 antibody
    • PDB 3 antibody
    • PDB3 antibody
    • Phosphotyrosine independent ligand for the Lck SH2 domain of 62 kDa antibody
    • Phosphotyrosine independent ligand for the Lck SH2 domain p62 antibody
    • Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa antibody
    • PKC-zeta-interacting protein antibody
    • Protein kinase C-zeta-interacting protein antibody
    • Sequestosome 1 antibody
    • Sequestosome-1 antibody
    • SQSTM 1 antibody
    • SQSTM_HUMAN antibody
    • Sqstm1 antibody
    • STAP antibody
    • STONE14 antibody
    • Ubiquitin binding protein p62 antibody
    • Ubiquitin-binding protein p62 antibody
    • ZIP 3 antibody
    • ZIP antibody
    • ZIP3 antibody
    see all

Images

  • All lanes : Anti-SQSTM1 / p62 antibody (ab155686) at 1/1000 dilution

    Lane 1 : Untreated (–) HepG2 whole cell extracts
    Lane 2 : HepG2 treated with 3µM Thapsigargin for 24 hours whole cell extracts (+)

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : Rabbit IgG antibody (HRP) at 1/10000 dilution

    Predicted band size: 47 kDa



    10% gel.

    Running condition: 80V, 15min; 140V, 40min.

    Transfer condition: Semi-dry, 18 V, 60min (Nitrocellulose membrane).

    Blocking condition: 5% non-fat milk in TBST, RT, 60min.

    Primary antibody incubation: 4°C overnight.

    Secondary antibody incubation: Room temperature for 1 hour.

    Washing condition: 5 ml TBST, 4 x 5min.

    ECL exposure. 

  • Immunocytochemistry/Immunofluorescent analysis of HeLa cells mock (left) and treated with 50μM Chloroquine for 24 hr (right) labeling SQSTM1/p62 (green) with ab155686 at 1/1000 dilution. Cells were fixed with 4% paraformaldehyde at RT for 15 min. Red: Phalloidin, a F-actin marker.

  • Immunoprecipitation of SQSTM1/p62 protein from HeLa whole cell extracts using 5 μg of ab155686.
    Western blot analysis was performed using ab155686.
    Anti-Rabbit IgG was used as a secondary reagent.

  • Paraffin-embedded human ovarian carcinoma tissue stained for SQSTM1/p62 with ab155686 (1/500) in Immunohistochemical analysis. Antigen retrieval: EDTA based buffer (pH 8) for 15 minutes. 

  • All lanes : Anti-SQSTM1 / p62 antibody (ab155686) at 1/500 dilution

    Lane 1 : HepG2 whole cell extracts
    Lane 2 : SQSTM1/p62 shRNA transfected HepG2 whole cell extracts

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG antibody

    Predicted band size: 47 kDa



    10% SDS-PAGE

  • 4% paraformaldehyde-fixed HeLa cells stained for SQSTM1/p62 (blue) using ab155686 (1/100) in Flow Cytometry. The cells were fixed at 4°C for 5 minutes. The brown line is unlabelled sample. Acquisition of >20,000 events were collected using Argon ion laser (488nm) and 525/30 bandpass filter.

  • Immunocytochemistry/ Immunofluorescence analysis of HepG2 cells treated with 3μM thapsigargin for 12 hours (right) and mock (left) labeling SQSTM1/p62 with ab155686 at 1:500 (green). Cells were fixed in ice-cold MeOH for 10 minutes, permeabilized with cooled acetone for 1 minute. Cells were co-stained with Hoechst (blue).

  • All lanes : Anti-SQSTM1 / p62 antibody (ab155686) at 1/1000 dilution

    Lane 1 : PC-12 whole cell lysate/extract
    Lane 2 : Rat2 whole cell lysate/extract

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-rabbit IgG antibody

    Predicted band size: 47 kDa



    10% SDS-PAGE

  • Anti-SQSTM1 / p62 antibody (ab155686) at 1/1000 dilution + A549 whole cell lysate at 30 µg

    Secondary
    HRP-conjugated anti-rabbit IgG antibody

    Predicted band size: 47 kDa



    10% SDS PAGE

References

This product has been referenced in:

  • Gong G  et al. Tanshinone IIA alleviates oxidative damage after spinal cord injury in vitro and in vivo through up-regulating miR-124. Life Sci 216:147-155 (2019). Read more (PubMed: 30468834) »
  • Ma Z  et al. Autophagy promotes hepatic differentiation of hepatic progenitor cells by regulating the Wnt/ß-catenin signaling pathway. J Mol Histol 50:75-90 (2019). Read more (PubMed: 30604254) »
See all 13 Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews

Application
Immunohistochemistry (Frozen sections)
Sample
Zebrafish Tissue sections (hindbrain at 8 somites)
Permeabilization
Yes - triton x-100 0.1%
Specification
hindbrain at 8 somites
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde

Adrian Santos

Verified customer

Submitted Nov 15 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (293T)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
293T
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 22 2018

Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (BV2)
Total protein in input
100 µg
Treatment
10 nM for 24hrs
Immuno-precipitation step
Protein A/G
Specification
BV2

Dr. jianhai long

Verified customer

Submitted Jan 15 2018

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