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SQSTM1 (p62) Antibody (C-term)

Purified Rabbit Polyclonal Antibody (Pab)

     
  • IF - SQSTM1 (p62) Antibody (C-term) AP2183B
    Fluorescent image of U251 cells stained with SQSTM1 (p62) (C-term) antibody. U251 cells were treated with Chloroquine (50 μM,16h), then fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with AP2183b SQSTM1 (p62) (C-term) primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min). SQSTM1 (p62) immunoreactivity is localized to autophagic vacuoles in the cytoplasm of U251 cells, supported by Human Protein Atlas Data (http://www.proteinatlas.org/ENSG00000161011).
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  • IF - SQSTM1 (p62) Antibody (C-term) AP2183B
    Immunofluorescence staining of Autophagy SQSTM1 (p62) Antibody (C-term) (Cat# AP2183b) on Methanol-fixed and PFA fixed HeLa cells. Data courtesy of Dr. Eeva-Liisa Eskelinen, University of Helsinki,Finland.
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  • WB - SQSTM1 (p62) Antibody (C-term) AP2183B
    All lanes : Anti-SQSTM1 (p62) Antibody (C-term) at 1:2000 dilution Lane 1: A549 whole cell lysate Lane 2: C2C12 whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 48 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
    detail
  • WB - SQSTM1 (p62) Antibody (C-term) AP2183B
    All lanes : Anti-SQSTM1 (p62) Antibody (C-term) at 1:2000 dilution Lane 1: Jurkat whole cell lysate Lane 2: THP-1 whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 48 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
    detail
  • IHC-P - SQSTM1 (p62) Antibody (C-term) AP2183B
    AP2183b staining SQSTM1 in Human prostate tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
    detail
  • IHC-P - SQSTM1 (p62) Antibody (C-term) AP2183B
    Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma.
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  • SPECIFICATION
  • CITATIONS: 30
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Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immunoelectronmicroscopy
  • EIA=Enzyme Immunoassay
WB, IF, IHC-P, E
Primary Accession Q13501
Other Accession O08623, Q64337
Reactivity Human, Mouse
Predicted Rat
Host Rabbit
Clonality Polyclonal
Isotype Rabbit IgG
Calculated MW 47687 Da
Antigen Region 317-346 aa
Additional Information
Gene ID 8878
Other Names Sequestosome-1, EBI3-associated protein of 60 kDa, EBIAP, p60, Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa, Ubiquitin-binding protein p62, SQSTM1, ORCA, OSIL
Target/Specificity This SQSTM1 (p62) antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 317-346 amino acids of human SQSTM1 (p62).
Dilution IF~~1:50~100
WB~~1:2000
IHC-P~~1:50~100
Format Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by peptide affinity purification.
StorageMaintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
PrecautionsSQSTM1 (p62) Antibody (C-term) is for research use only and not for use in diagnostic or therapeutic procedures.
Protein Information
Name SQSTM1
Synonyms ORCA, OSIL
Function Autophagy receptor required for selective macroautophagy (aggrephagy) (PubMed:34471133, PubMed:16286508, PubMed:20168092, PubMed:24128730, PubMed:28404643, PubMed:22622177, PubMed:33509017). Functions as a bridge between polyubiquitinated cargo and autophagosomes (PubMed:34471133). Interacts directly with both the cargo to become degraded and an autophagy modifier of the MAP1 LC3 family (PubMed:16286508, PubMed:20168092, PubMed:24128730, PubMed:28404643, PubMed:22622177). Along with WDFY3, involved in the formation and autophagic degradation of cytoplasmic ubiquitin- containing inclusions (p62 bodies, ALIS/aggresome-like induced structures). Along with WDFY3, required to recruit ubiquitinated proteins to PML bodies in the nucleus (PubMed:24128730, PubMed:20168092). Also involved in autophagy of peroxisomes (pexophagy) in response to reactive oxygen species (ROS) by acting as a bridge between ubiquitinated PEX5 receptor and autophagosomes (PubMed:26344566). 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. Involved in endosome organization by retaining vesicles in the perinuclear cloud: following ubiquitination by RNF26, attracts specific vesicle-associated adapters, forming a molecular bridge that restrains cognate vesicles in the perinuclear region and organizes the endosomal pathway for efficient cargo transport (PubMed:27368102). Promotes relocalization of 'Lys-63'-linked ubiquitinated STING1 to autophagosomes (PubMed:29496741). Acts as an activator of the NFE2L2/NRF2 pathway via interaction with KEAP1: interaction inactivates the BCR(KEAP1) complex, promoting nuclear accumulation of NFE2L2/NRF2 and subsequent expression of cytoprotective genes (PubMed:20452972, PubMed:28380357, PubMed:33393215). Sequesters tensin TNS2 into cytoplasmic puncta, promoting TNS2 ubiquitination and proteasomal degradation (PubMed:25101860).
Cellular Location Cytoplasm, cytosol. Preautophagosomal structure. Late endosome. Lysosome. Cytoplasmic vesicle, autophagosome. Nucleus. Endoplasmic reticulum. Nucleus, PML body. Cytoplasm, myofibril, sarcomere. Note=In cardiac muscle, localizes to the sarcomeric band (By similarity). Commonly found in inclusion bodies containing polyubiquitinated protein aggregates. In neurodegenerative diseases, detected in Lewy bodies in Parkinson disease, neurofibrillary tangles in Alzheimer disease, and HTT aggregates in Huntington disease. In protein aggregate diseases of the liver, found in large amounts in Mallory bodies of alcoholic and nonalcoholic steatohepatitis, hyaline bodies in hepatocellular carcinoma, and in SERPINA1 aggregates Enriched in Rosenthal fibers of pilocytic astrocytoma. In the cytoplasm, observed in both membrane-free ubiquitin-containing protein aggregates (sequestosomes) and membrane-surrounded autophagosomes Colocalizes with TRIM13 in the perinuclear endoplasmic reticulum. Co- localizes with TRIM5 in cytoplasmic bodies. When nuclear export is blocked by treatment with leptomycin B, accumulates in PML bodies
Tissue Location Ubiquitously expressed.
Research Areas
Citations ( 0 )

Background

SQSTM1/p62 is an adapter protein which binds ubiquitin and may regulate the activation of NFKB1 by TNF-alpha, nerve growth factor (NGF) and interleukin-1. This protein may play a role in titin/TTN downstream signaling in muscle cells, and may also regulate signaling cascades through ubiquitination. This protein is involved in cell differentiation, apoptosis, immune response and regulation of K(+) channels. SQSTM1/p62 also appears to play a role in macroautophagic removal of intracellular protein aggregates. Cellular depletion studies of SQSTM1/p62 have indicated a role for association with LC3 and aggregate proteins in order to facilitate normal formation of the autophagosome.

References

References for protein:
1.Seibenhener, M.L., et al., Mol. Cell. Biol. 24(18):8055-8068 (2004).
2.Eekhoff, E.W., et al., Arthritis Rheum. 50(5):1650-1654 (2004).
3.Brajenovic, M., et al., J. Biol. Chem. 279(13):12804-12811 (2004).
4.Kuusisto, E., et al., J. Neuropathol. Exp. Neurol. 62(12):1241-1253 (2003).
5. Johnson-Pais, T.L., et al., J. Bone Miner. Res. 18(10):1748-1753 (2003).
References for U251 cell line:
1. Westermark B.; Pontén J.; Hugosson R. (1973).” Determinants for the establishment of permanent tissue culture lines from human gliomas”. Acta Pathol Microbiol Scand A. 81:791-805. [PMID: 4359449].
2. Pontén, J.,Westermark B. (1978).” Properties of Human Malignant Glioma Cells in Vitro”. Medical Biology 56: 184-193.[PMID: 359950].
3. Geng Y.;Kohli L.; Klocke B.J.; Roth K.A.(2010). “Chloroquine-induced autophagic vacuole accumulation and cell death in glioma cells is p53 independent”. Neuro Oncol. 12(5): 473–481.[ PMID: 20406898].

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Cat# AP2183B
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