Human/Mouse/Rat p62/SQSTM1 Antibody

Catalog # Availability Size / Price Qty
MAB8028
MAB8028-SP
Detection of Human, Mouse, and Rat p62/SQSTM1 by Western Blot.
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Product Details
Citations (18)
FAQs
Supplemental Products
Reviews (2)

Human/Mouse/Rat p62/SQSTM1 Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human p62/SQSTM1 in ELISAs. Detects human, mouse and rat p62/SQSTM1 in Western blots
Source
Monoclonal Mouse IgG1 Clone # 864807
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
E. coli-derived recombinant human p62/SQSTM1
Asp368-Leu440
Accession # Q13501
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 µg/mL
See below
Simple Western
20 µg/mL
See below
Immunohistochemistry
5-25 µg/mL
Immersion fixed paraffin-embedded sections of human liver
Immunoprecipitation
1 µg/1 mg cell lysate
Cell lysate of U2OS human osteosarcoma cell line 
Immunocytochemistry
8-25 µg/mL
See below
Knockout Validated
p62/SQSTM1 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line and parental U2OS cell line but is not detectable in p62/SQSTM1 knockout HeLa cell line and knockout U2OS cell line. 
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human, Mouse, and Rat p62/SQSTM1 antibody by Western Blot. View Larger

Detection of Human, Mouse, and Rat p62/SQSTM1 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, RAW 264.7 mouse monocyte/macrophage cell line, and C6 rat glioma cell line untreated (-) or treated (+) with 1 µg/mL LPS for 24 hours. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for p62/SQSTM1 at approximately 62 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry p62/SQSTM1 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC). View Larger

p62/SQSTM1 in HeLa Human Cell Line. p62/SQSTM1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to phagosomes in cell cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry View Larger

p62/SQSTM1 in Human Liver. p62/SQSTM1 was detected in immersion fixed paraffin-embedded sections of human liver using Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei and cytoplasm in hepatocytes. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western Detection of Mouse p62/SQSTM1 antibody by Simple Western<SUP>TM</SUP>. View Larger

Detection of Mouse p62/SQSTM1 by Simple WesternTM. Simple Western lane view shows lysates of RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with 1 µg/mL LPS for 24 hours, loaded at 0.2 mg/mL. A specific band was detected for p62/SQSTM1 at approximately 66 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Knockout Validated p62/SQSTM1 Antibody Specificity is Shown by Immunocytochemistry antibody in Knockout Cell Line. View Larger

p62/SQSTM1 Specificity is Shown by Immunocytochemistry in Knockout Cell Line. p62/SQSTM1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line but is not detected in p62/SQSTM1 knockout (KO) HeLa cell line using Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Knockout Validated Western Blot Shows Human p62/SQSTM1 Antibody Specificity Using Knockout Cell Line. View Larger

Western Blot Shows Human p62/SQSTM1 Specificity Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and p62/SQSTM1 knockout HeLa cell line (KO). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for p62/SQSTM1 at approximately 62 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. GAPDH (MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Knockout Validated View Larger

Western Blot Shows Human p62/SQSTM1 Specificity Using Knockout Cell Line. Western blot shows lysates of U2OS human osteosarcoma cell line and p62/SQSTM1 knockout U2OS cell line (KO). Nitrocellulose membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat p62/SQSTM1 Monoclonal Antibody (Catalog # MAB8028) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody. A specific band was detected for p62/SQSTM1 at approximately 62 kDa (as indicated) in the parental U2OS cell line, but is not detectable in knockout U2OS cell line. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

Immunoprecipitation View Larger

Detection of SQSTM1/p62 by Immunoprecipitation Immunoprecipitation was performed on cell lysate of U2OS human osteosarcoma cell line using 1.0 μg of Mouse Anti-Human SQSTM1/p62 Monoclonal Antibody (Catalog # MAB8028) pre-coupled to protein G or protein A beads. Immunoprecipitated SQSTM1/p62 was detected with Rabbit Anti-SQSTM1/p62 Monoclonal Antibody (Catalog # MAB80281). The Ponceau stained transfers of each blot are shown. SM=10% starting material; UB=10% unbound fraction; IP=immunoprecipitated. Image, protocol, and testing courtesy of YCharOS Inc. (ycharos.com).

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.5 mg/mL.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: p62/SQSTM1

SQSTM1 (Sequestrome-1), also called p62, is a widely expressed, stress-inducible, multifunctional 62 kDa intracellular protein. The 440 amino acid (aa) human SQSTM1 contains multiple adaptor domains that allow interaction with proteins in NGF/NFkB and other signaling pathways (notably TRAF6, atypical protein kinase C family and Src family), polyubiquitin, proteasome subunits and many others. It contains numerous regulatory phosphorylation sites and a dimerization site. SQSTM1 shuttles ubiquitinylated proteins to the proteasome and is important in autophagy and apoptosis. Its dysregulation is associated with Paget’s disease of bone, Parkinson’s and Alzheimer’s diseases, and cancers. Within aa 344-440, which includes the ubiquitin-binding domain, human SQSTM1 shares 100% aa sequence identity with mouse and rat SQSTM1.

Long Name
Sequestosome 1
Entrez Gene IDs
8878 (Human); 18412 (Mouse); 113894 (Rat)
Alternate Names
A170; Autophagy Receptor P62; DMRV; EBI3-associated protein of 60 kDa; EBI3-associated protein p60; EBIAP; FTDALS3; NADGP; ORCA; OSIL; oxidative stress induced like; p60PDB3; p62; p62B; Paget disease of bone 3; PDB3; phosphotyrosine independent ligand for the Lck SH2 domain p62; Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa; Sequestosome 1; sequestosome-1; SQSTM1; Ubiquitin-binding protein p62; ZIP3

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Citations for Human/Mouse/Rat p62/SQSTM1 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

18 Citations: Showing 1 - 10
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  1. Lenalidomide increases human dendritic cell maturation in multiple myeloma patients targeting monocyte differentiation and modulating mesenchymal stromal cell inhibitory properties
    Authors: Costa F, Vescovini R, Bolzoni M et al.
    Oncotarget
  2. mtDNA variability determines spontaneous joint aging damage in a conplastic mouse model
    Authors: Morena Scotece, Carlos Vaamonde-García, Ana Victoria Lechuga-Vieco, Alberto Centeno Cortés, María Concepción Jiménez Gómez, Purificación Filgueira-Fernández et al.
    Aging (Albany NY)
  3. Increased expression of toll-like receptor 3, an anti-viral signaling molecule, and related genes in Alzheimer's disease brains.
    Authors: Walker D, Tang T, Lue L
    Exp Neurol, 2018-08-01;309(0):91-106.
  4. Histone H2A ubiquitination resulting from Brap loss of function connects multiple aging hallmarks and accelerates neurodegeneration
    Authors: Guo Y, Chomiak AA, Hong Y et al.
    iScience
  5. Sex and age differences in AMPK phosphorylation, mitochondrial homeostasis, and inflammation in hearts from inflammatory cardiomyopathy patients
    Authors: Barcena ML, Tonini G, Haritonow N et al.
    Aging cell
  6. Junín virus induces autophagy in human A549 cells
    Authors: Maria Laura A. Perez Vidakovics, Agustín E. Ure, Paula N. Arrías, Víctor Romanowski, Ricardo M. Gómez
    PLOS ONE
  7. Absence of either Ripk3 or Mlkl reduces incidence of hepatocellular carcinoma independent of liver fibrosis
    Authors: Mohammed S, Thadathil N, Ohene-Marfo P et al.
    Molecular cancer research : MCR
  8. The identification of high-performing antibodies for Sequestosome-1 for use in Western blot, immunoprecipitation and immunofluorescence
    Authors: Ayoubi, R;Alshafie, W;Shlaifer, I;Southern, K;McPherson, PS;Laflamme, C;NeuroSGC/YCharOS/EDDU collaborative group, ;
    F1000Research
    Species: Human
    Sample Types: Cell Lysates, Whole Cells, Transfected Whole Cells
    Applications: Immunoprecipitation, Western Blot, Immunocytochemistry
  9. Treprostinil Reconstitutes Mitochondrial Organisation and Structure in Idiopathic Pulmonary Fibrosis Cells
    Authors: Fang, L;Chen, WC;Jaksch, P;Molino, A;Saglia, A;Roth, M;Lambers, C;
    International journal of molecular sciences
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  10. Role of macrophage autophagy in postoperative pain and inflammation in mice
    Authors: Mitsui, K;Hishiyama, S;Jain, A;Kotoda, Y;Abe, M;Matsukawa, T;Kotoda, M;
    Journal of neuroinflammation
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  11. Airway Smooth Muscle Cell Mitochondria Damage and Mitophagy in COPD via ERK1/2 MAPK
    Authors: L Fang, M Zhang, J Li, L Zhou, M Tamm, M Roth
    International Journal of Molecular Sciences, 2022-11-12;23(22):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  12. Inhibition of Carbonic Anhydrase 2 Overcomes Temozolomide Resistance in Glioblastoma Cells
    Authors: K Zhao, A Schäfer, Z Zhang, K Elsässer, C Culmsee, L Zhong, A Pagenstech, C Nimsky, JW Bartsch
    International Journal of Molecular Sciences, 2021-12-23;23(1):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  13. Rapid 3D phenotypic analysis of neurons and organoids using data-driven cell segmentation-free machine learning
    Authors: P Mergenthal, S Hariharan, JM Pemberton, C Lourenco, LZ Penn, DW Andrews
    PLoS computational biology, 2021-02-22;17(2):e1008630.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  14. Wnt/&beta-catenin signaling pathway induces autophagy-mediated temozolomide-resistance in human glioblastoma
    Authors: EJ Yun, S Kim, JT Hsieh, ST Baek
    Cell Death Dis, 2020-09-17;11(9):771.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  15. Cold-inducible RNA-binding protein through TLR4 signaling induces mitochondrial DNA fragmentation and regulates macrophage cell death after trauma
    Authors: Z Li, EK Fan, J Liu, MJ Scott, Y Li, S Li, W Xie, TR Billiar, MA Wilson, Y Jiang, P Wang, J Fan
    Cell Death Dis, 2017-05-11;8(5):e2775.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  16. Tribbles Pseudokinase 3 Induces Both Apoptosis and Autophagy in Amyloid-? induced Neuronal Death
    Authors: Suraiya Saleem
    J. Biol. Chem, 2016-12-23;0(0):.
    Species: Mouse, Rat
    Sample Types: Cell Lysates, Whole Cells
    Applications: ICC, Western Blot
  17. Preventing mutant huntingtin proteolysis and intermittent fasting promote autophagy in models of Huntington disease
    Authors: Dagmar E. Ehrnhoefer, Dale D. O. Martin, Mandi E. Schmidt, Xiaofan Qiu, Safia Ladha, Nicholas S. Caron et al.
    Acta Neuropathologica Communications
  18. Defective Lipid Droplet–Lysosome Interaction Causes Fatty Liver Disease as Evidenced by Human Mutations in TMEM199 and CCDC115
    Authors: Lars E. Larsen, Marjolein A.W. van den Boogert, Wilson A. Rios-Ocampo, Jos C. Jansen, Donna Conlon, Patrick L.E. Chong et al.
    Cellular and Molecular Gastroenterology and Hepatology

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Reviews for Human/Mouse/Rat p62/SQSTM1 Antibody

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Human/Mouse/Rat p62/SQSTM1 Antibody
By Anonymous on 08/15/2021
Application: Immunocytochemistry/Immunofluorescence Sample Tested: HeLa cells Species: Human

Human/Mouse/Rat p62/SQSTM1 Antibody
By Anonymous on 10/17/2018
Application: WB Sample Tested: HeLa cells Species: Human