Human/Mouse/Rat p62/SQSTM1 Antibody

(1 citations)   
  • 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 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
  • Immunocytochemistry
    8-25 µg/mL
    See below
  • Knockout Validated
    p62/SQSTM1 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in p62/SQSTM1 knockout HeLa 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.
Data Examples
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 (Catalog # 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 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; Catalog # 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.
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 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; Catalog # 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 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 (Catalog # 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 knockoutHeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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; EBI3-associated protein of 60 kDa; EBI3-associated protein p60; EBIAP; 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
Related Research Areas
Citations:

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.

1 Citations: Showing 1 - 1

  1. 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;8(5):e2775.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
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