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    sqstm1--p62-antibody-epr18351-ab207305.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (3) Submit a question References (25)

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Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
  • Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18351] to SQSTM1 / p62
  • Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IF, IP
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-SQSTM1 / p62 antibody [EPR18351]
    See all SQSTM1 / p62 primary antibodies
  • Description

    Rabbit monoclonal [EPR18351] to SQSTM1 / p62
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human liver and kidney lysates; U-2 OS, HCT116, HepG2 and HeLa whole cell lysates. IHC-P: Human hepatocellular carcinoma and lung cancer tissues. ICC/IF: HeLa cells (untreated and treated with chloroquine), HAP1 cells (untreated and treated with chloroquine), U-2 OS cells. Flow Cyt (intra): PC-3 cells. IP: HeLa and HepG2 whole cell lysates.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18351
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFkB Pathway
    • Signal Transduction
    • Protein Trafficking
    • Vesicle Transport
    • Regulation
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Polymerase associated factors
    • Pol II Transcription
    • Other
    • Cardiovascular
    • Heart
    • Autophagy
    • Autophagosome
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • Autophagosome
    • Cancer
    • Cell Death
    • Autophagy
    • Autophagosome
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-SQSTM1 / p62 antibody [EPR18351] (ab225453)
    • PE Anti-SQSTM1 / p62 antibody [EPR18351] (ab225454)
    • Anti-SQSTM1 / p62 antibody [EPR18351] - BSA and Azide free (ab227992)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human SQSTM1 (p62) knockout HCT116 cell line (ab266871)
  • KO cell lysates

    • Human SQSTM1 (p62) knockout HCT116 cell lysate (ab257052)
  • Recombinant Protein

    • Recombinant Human SQSTM1 / p62 protein (ab95320)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab207305 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
Flow Cyt (Intra)
1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 47 kDa).
ICC/IF (1)
Use a concentration of 1 µg/ml.
IP (2)
1/40.
Notes
Flow Cyt (Intra)
1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 47 kDa).
ICC/IF
Use a concentration of 1 µg/ml.
IP
1/40.

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.
  • Target information above from: UniProt accession Q13501 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8878 Human
    • Omim: 601530 Human
    • SwissProt: Q13501 Human
    • Unigene: 709030 Human
    • 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

    • Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: SQSTM1/p62 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: HepG2 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green).

      Green - target observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.

      This western blot image is a comparison between ab207305 and a competitor's discontinued goat polyclonal antibody.

       

       

    • Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      ab207305 was shown to react with SQSTM1 in wild-type U-2 OS cells in Immunocytochemistry with loss of signal observed in a SQSTM1 knockout cell line. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1/5000. The cells were then incubated with ab207305 at 1/200 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
    • Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      All lanes : Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

      Lane 1 : Wild-type HCT116 cell lysate
      Lane 2 : SQSTM1 CRISPR/Cas9 edited HCT116 cell lysate
      Lane 3 : HepG2 cell lysate
      Lane 4 : HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 47 kDa
      Observed band size: 55 kDa why is the actual band size different from the predicted?



      Lanes 1-4: Merged signal (red and green). Green - ab207305 observed at 55 kDa. Red - loading control ab8245 observed at 36 kDa.

       ab207305 Anti-SQSTM1 / p62 antibody [EPR18351] was shown to specifically react with SQSTM1 / p62 in wild-type HCT116 cells. The band observed in CRISPR/Cas9 edited cell line ab266871 (CRISPR/Cas9 edited cell lysate ab257052) lane below 55 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and SQSTM1 / p62 CRISPR/Cas9 edited samples were subjected to SDS-PAGE. ab207305 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

      ab207305 staining SQSTM1 in wild-type HAP1 cells and knockout cells, untreated and chloroquine-treated (ab142116, 50μM, 24 hours). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab207305 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      All lanes : Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305) at 1/1000 dilution

      Lane 1 : Wild-type U-2 OS cell lysate
      Lane 2 : SQSTM1 knockout U-2 OS cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 47 kDa



      ab207305 was shown to react with SQSTM1 in wild-type U-2 OS cells in Western blot with loss of signal observed in a SQSTM1 knockout cell line. Wild-type U-2 OS and SQSTM1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab207305 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2ug/mL before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

    • Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: SQSTM1/p62 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: HepG2 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab207305 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab207305 was shown to specifically react with SQSTM1/p62 in wild-type HAP1 cells. No band was observed when SQSTM1/p62 knockout samples were used. Wild-type and SQSTM1/p62 knockout samples were subjected to SDS-PAGE, ab207305 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

      Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling SQSTM1 / p62 with ab207305 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on human hepatocellular carcinoma is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

      ab207305 staining SQSTM1/p62 in HeLa cells +/- Chloroquine (50μM, 24 hours). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab207305 at 1μg/ml and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in pseudocolor red) followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      All lanes : Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305) at 1/1000 dilution

      Lane 1 : Human liver lysate
      Lane 2 : Human kidney lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

      Predicted band size: 47 kDa
      Observed band size: 62 kDa why is the actual band size different from the predicted?


      Exposure time: 30 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The MW observed is consistent with what has been described in the literature (PMID: PMC4344198).

    • Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      All lanes : Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305) at 1/5000 dilution

      Lane 1 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
      Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 47 kDa
      Observed band size: 62 kDa why is the actual band size different from the predicted?


      Exposure time: 30 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The MW observed is consistent with what has been described in the literature (PMID: PMC4344198).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

      Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling SQSTM1 / p62 with ab207305 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm and weakly nucleus staining on human lung cancer is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

      Immunohistochemical analysis of paraffin-embedded human liver tissue labeling SQSTM1 / p62 with ab207305 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Hhman normal liver tissue. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed PC-3 (Human prostate adenocarcinoma cell line) cells labeling SQSTM1 / p62 with ab207305 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody. 

       

       

    • Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

      SQSTM1 / p62 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab207305 at 1/40 dilution.
      Western blot was performed from the immunoprecipitate using ab207305 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
      Lane 1: HeLa whole cell lysate 10ug (Input).
      Lane 2: ab207305 IP in HeLa whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207305 in HeLa whole cell lysate.
      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 1 second.

    • Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

      SQSTM1 / p62 was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell lysate with ab207305 at 1/40 dilution.
      Western blot was performed from the immunoprecipitate using ab207305 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
      Lane 1: HepG2 whole cell lysate 10ug (Input).
      Lane 2: ab207305 IP in HepG2 whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207305 in HepG2 whole cell lysate.
      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 1 second.

    • Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)
      Anti-SQSTM1 / p62 antibody [EPR18351] (ab207305)

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

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    References (25)

    Publishing research using ab207305? Please let us know so that we can cite the reference in this datasheet.

    ab207305 has been referenced in 25 publications.

    • Zhang L  et al. S100A8 promotes chemoresistance via augmenting autophagy in B-cell lymphoma cells. Oncol Rep 45:151-158 (2021). PubMed: 33169801
    • Xu W  et al. YAP manipulates proliferation via PTEN/AKT/mTOR-mediated autophagy in lung adenocarcinomas. Cancer Cell Int 21:30 (2021). PubMed: 33413409
    • Wang J  et al. Autophagy augments the self-renewal of lung cancer stem cells by the degradation of ubiquitinated p53. Cell Death Dis 12:98 (2021). PubMed: 33468994
    • Hickman RA  et al. Developmental malformations in Huntington disease: neuropathologic evidence of focal neuronal migration defects in a subset of adult brains. Acta Neuropathol 141:399-413 (2021). PubMed: 33517535
    • Chen Y  et al. Epidemiology of Drug- and Herb-Induced Liver Injury Assessed for Causality Using the Updated RUCAM in Two Hospitals from China. Biomed Res Int 2021:8894498 (2021). PubMed: 33954202
    View all Publications for this product

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Immunoprecipitation abreview for Anti-SQSTM1 / p62 antibody [EPR18351]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (human kidney)
    Total protein in input
    200 µg
    Immuno-precipitation step
    Protein A
    Specification
    human kidney
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Sep 06 2018

    Immunoprecipitation abreview for Anti-SQSTM1 / p62 antibody [EPR18351]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Mouse Cell lysate - whole cell (Mouse Embyronic fibroblasts)
    Total protein in input
    200 µg
    Immuno-precipitation step
    Protein A
    Specification
    Mouse Embyronic fibroblasts
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 22 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-SQSTM1 / p62 antibody [EPR18351]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Mouse Embyronic fibroblasts)
    Permeabilization
    Yes - Digitonin
    Specification
    Mouse Embyronic fibroblasts
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Dr. Ryan Russell

    Verified customer

    Submitted Aug 10 2017

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