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    products/primary-antibodies/sqstm1--p62-phospho-s349-antibody-epr20451-ab211324.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (3) Submit a question References (10)

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Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
  • Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
  • Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
  • Immunoprecipitation - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
  • Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
  • Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
  • Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
  • Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20451] to SQSTM1 / p62 (phospho S349)
  • Suitable for: Flow Cyt (Intra), ICC/IF, IP, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

You may also be interested in

Primary
Product image
Alexa Fluor® 488 Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab237322)
Knockout
Product image
Human SQSTM1 (p62) knockout HEK-293T cell line (ab255343)
Protein
Product image
Recombinant Human SQSTM1 / p62 protein (ab95320)

View more associated products

Overview

  • Product name

    Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451]
    See all SQSTM1 / p62 primary antibodies
  • Description

    Rabbit monoclonal [EPR20451] to SQSTM1 / p62 (phospho S349)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: U-2 OS, HeLa, C6 and NIH/3T3 whole cell lysates treated with 2µM MG-132 (ab141003) for 18 hours. ICC/IF: HeLa cells treated with 2µM MG-132 (ab141003) for 18 hours. Flow Cyt (intra): HeLa cells treated with 2µM MG-132 (ab141003) for 18 hours. IP: HeLa treated with 2µM MG-132 (ab141003) for 18h whole cell lysate.
  • 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, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20451
  • 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® 488 Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab237322)
    • Alexa Fluor® 647 Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab237323)
    • Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] - BSA and Azide free (ab238445)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

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

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • MG-132, proteasome inhibitor (ab141003)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab211324 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)
1/500.
ICC/IF (1)
1/100.
IP
1/30.
WB (2)
1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 47 kDa).
Notes
Flow Cyt (Intra)
1/500.
ICC/IF
1/100.
IP
1/30.
WB
1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 47 kDa).

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
    • Entrez Gene: 18412 Mouse
    • Entrez Gene: 113894 Rat
    • Omim: 601530 Human
    • SwissProt: Q13501 Human
    • SwissProt: Q64337 Mouse
    • SwissProt: O08623 Rat
    • Unigene: 709030 Human
    • Unigene: 40828 Mouse
    • Unigene: 107103 Rat
    see all
  • 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 (phospho S349) antibody [EPR20451] (ab211324)
    Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
    All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) 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



    ab211324 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 ab211324 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
    Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 2μM MG-132 (ab141003) for 18 hours or untreated, labeling SQSTM1 / p62 (phospho S349) with ab211324 at 1/100 dilution, followed by Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) secondary antibody at 1/200 dilution (green).

    Confocal image showing cytoplasmic staining on HeLa cell line. The expression increased after treatment with 2μM MG-132 (ab141003) for 18 hours.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
    Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 2µM MG-132 (ab141003) for 18 hours (red) or untreated (green), labeling SQSTM1 / p62 (phospho S349) with ab211324 at 1/500 dilution compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody. 

     

     

  • Immunoprecipitation - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
    Immunoprecipitation - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

    SQSTM1 / p62 (phospho S349) was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 2μM MG-132 (ab141003) for 18h with ab211324 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab211324 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Lane 1: HeLa treated with 2μM MG-132 (ab141003) for 18h whole cell lysate, 10 µg (Input).
    Lane 2: ab211324 IP in HeLa treated with 2μM MG-132 (ab141003) for 18h whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab211324 in HeLa treated with 2μM MG-132 (ab141003) for 18h whole cell lysate.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
    Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
    All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution

    Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours
    Lane 3 : HeLa whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours, then treated with Alkaline Phosphatase for 1 hour

    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: 1 minute


    Blocking/dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
    Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
    All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution

    Lane 1 : Untreated C6 (Rat glial tumor cell line) whole cell lysate
    Lane 2 : C6 whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours

    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: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
    Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
    All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution

    Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 2 : NIH/3T3 whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours

    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: 4 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
    Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)

Protocols

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

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (10)

Publishing research using ab211324? Please let us know so that we can cite the reference in this datasheet.

ab211324 has been referenced in 10 publications.

  • Shen Z  et al. Inhibition of HDAC6 by Tubastatin A reduces chondrocyte oxidative stress in chondrocytes and ameliorates mouse osteoarthritis by activating autophagy. Aging (Albany NY) 13:9820-9837 (2021). PubMed: 33744850
  • Chen TT  et al. Gastrodin ameliorates learning and memory impairment in rats with vascular dementia by promoting autophagy flux via inhibition of the Ca2+/CaMKII signal pathway. Aging (Albany NY) 13:9542-9565 (2021). PubMed: 33714957
  • Li M  et al. Autophagy in the HTR-8/SVneo Cell Oxidative Stress Model Is Associated with the NLRP1 Inflammasome. Oxid Med Cell Longev 2021:2353504 (2021). PubMed: 33854691
  • Xu Q  et al. Adenosine 5'-monophosphate-activated protein kinase ameliorates bovine adipocyte oxidative stress by inducing antioxidant responses and autophagy. J Dairy Sci 104:4516-4528 (2021). PubMed: 33551161
  • Guo J  et al. Bone Marrow-Derived Mesenchymal Stem Cells Ameliorate Sepsis-Induced Acute Kidney Injury by Promoting Mitophagy of Renal Tubular Epithelial Cells via the SIRT1/Parkin Axis. Front Endocrinol (Lausanne) 12:639165 (2021). PubMed: 34248837
  • Gallego-Selles A  et al. Regulation of Nrf2/Keap1 signalling in human skeletal muscle during exercise to exhaustion in normoxia, severe acute hypoxia and post-exercise ischaemia: Influence of metabolite accumulation and oxygenation. Redox Biol 36:101627 (2020). PubMed: 32863217
  • Xu Q  et al. Cardamonin Reduces Acetaminophen-Induced Acute Liver Injury in Mice via Activating Autophagy and NFE2L2 Signaling. Front Pharmacol 11:601716 (2020). PubMed: 33364966
  • Shrishrimal S  et al. Manganese porphyrin, MnTE-2-PyP, treatment protects the prostate from radiation-induced fibrosis (RIF) by activating the NRF2 signaling pathway and enhancing SOD2 and sirtuin activity. Free Radic Biol Med 152:255-270 (2020). PubMed: 32222469
  • Gao Y  et al. Role of Parkin-mediated mitophagy in the protective effect of polydatin in sepsis-induced acute kidney injury. J Transl Med 18:114 (2020). PubMed: 32131850
  • Kerins MJ  et al. Genome-Wide CRISPR Screen Reveals Autophagy Disruption as the Convergence Mechanism That Regulates the NRF2 Transcription Factor. Mol Cell Biol 39:N/A (2019). PubMed: 31010806

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Western blot abreview for Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Non-Denaturing (Native) (10-12.5% polyacrilamide gel Wet transfer at 400mA for 90 min.)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: RT°C
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.

Marcos Martin Rincon

Verified customer

Submitted Sep 01 2022

Immunocytochemistry/ Immunofluorescence abreview for Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (C42B Prostate cancer cell line)
Permeabilization
Yes - 0.1% Triton X-100
Specification
C42B Prostate cancer cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
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. Dimitra Kalamida

Verified customer

Submitted Feb 28 2020

Western blot abreview for Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (C42B Prostate cancer cell line)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Specification
C42B Prostate cancer cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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. Dimitra Kalamida

Verified customer

Submitted May 14 2019

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