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Human SQSTM1 (p62) knockout HEK-293T cell lysate (ab263770)

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Western blot - Human SQSTM1 (p62) knockout HEK293T cell lysate (ab263770)
  • Western blot - Human SQSTM1 (p62) knockout HEK293T cell lysate (ab263770)
  • Sanger Sequencing - Human SQSTM1 knockout HEK293T cell lysate (ab263770)

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Protein
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Recombinant Human SQSTM1 / p62 protein (ab95320)

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Overview

  • Product name

    Human SQSTM1 (p62) knockout HEK-293T cell lysate
    See all SQSTM1 / p62 kits
  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    HEK293T
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 4.
  • Passage number

    <20
  • Knockout validation

    Sanger Sequencing, Western Blot (WB)
  • Reconstitution notes

    To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.

    *Usage of SDS sample buffer is not recommended with these lyophilized lysates.

  • Notes

    Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.

    User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

    Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
    See here for more information on knockout cell lysates.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

    This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab255452 - Human SQSTM1 knockout HEK293T cell lysate 1 x 100µg
    ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg
  • 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
  • Cell type

    epithelial
  • STR Analysis

    Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12

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
  • Alternative names

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

Associated products

  • KO cell lines

    • Human SQSTM1 (p62) knockout HEK-293T cell line (ab255343)
  • Related Products

    • Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker (ab109012)
    • Anti-SQSTM1 / p62 antibody [EPR4844] - BSA and Azide free (ab219581)
    • Anti-SQSTM1 / p62 antibody [2C11] - BSA and Azide free (ab56416)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab263770 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
WB
Use at an assay dependent concentration.
Notes
WB
Use at an assay dependent concentration.

Images

  • Western blot - Human SQSTM1 (p62) knockout HEK293T cell lysate (ab263770)
    Western blot - Human SQSTM1 (p62) knockout HEK293T cell lysate (ab263770)

    Lane 1: Wild-type HEK293T cell lysate (20µg)
    Lane 2: SQSTM1 knockout HEK293T cell lysate (20µg)
    Lanes 1- 2: Merged signal (red and green). Green - ab56416 observed at 62 kDa. Red - loading control ab181602 observed at 37 kDa.
    ab56416 Anti-SQSTM1 / p62 antibody - Autophagosome Marker was shown to specifically react with SQSTM1 in wild-type HEK293T cells in western blot. Loss of signal was observed when knockout cell line ab255343 (knockout cell lysate ab263770) was used. Wild-type and SQSTM1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab56416 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Human SQSTM1 (p62) knockout HEK293T cell lysate (ab263770)
    Western blot - Human SQSTM1 (p62) knockout HEK293T cell lysate (ab263770)

    Lane 1: Wild-type HEK293T cell lysate (20µg)
    Lane 2: SQSTM1 knockout HEK293T cell lysate (20µg)
    Lanes 1- 2: Merged signal (red and green). Green - ab109012 observed at 64 kDa. Red - loading control ab8245 observed at 37 kDa.
    ab109012 Recombinant Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker was shown to specifically react with SQSTM1 in wild-type HEK293T cells in western blot. Loss of signal was observed when knockout cell line ab255343 (knockout cell lysate ab263770) was used. Wild-type and SQSTM1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109012 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 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.

  • Sanger Sequencing - Human SQSTM1 knockout HEK293T cell lysate (ab263770)
    Sanger Sequencing - Human SQSTM1 knockout HEK293T cell lysate (ab263770)
    Homozygous: 1 bp insertion in exon 4

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab263770? Please let us know so that we can cite the reference in this datasheet.

ab263770 has not yet been referenced specifically in any publications.

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