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Signal Transduction Signaling Pathway Nuclear Signaling NFkB Pathway
使用敲除细胞株进行验证

Anti-SQSTM1 / p62抗体- Autophagosome Marker (ab56416)

  • Datasheet
  • SDS
Reviews (35)Q&A (4)References (499)

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Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)
  • Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)
  • Flow Cytometry - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)

Key features and details

  • Mouse monoclonal to SQSTM1 / p62 - Autophagosome Marker
  • Suitable for: IHC-P, WB, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG2a

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概述

  • 产品名称

    Anti-SQSTM1 / p62抗体- Autophagosome Marker
    参阅全部 SQSTM1 / p62 一抗
  • 描述

    小鼠单克隆抗体to SQSTM1 / p62 - Autophagosome Marker
  • 宿主

    Mouse
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • 免疫原

    Recombinant full length protein corresponding to Human SQSTM1/ p62 aa 1-440.
    Database link: Q13501

  • 阳性对照

    • WB: HeLa, Hap1. Flow Cyt: HeLa. IHC-P: Human lymph node. ICC/IF: HeLa
  • 常规说明

    This product was changed from ascites to tissue culture supernatant on 28th May 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

     

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液

    pH: 7.40
    Constituent: PBS
  • Concentration information loading...
  • 纯度

    Tissue culture supernatant
  • 克隆

    单克隆
  • 同种型

    IgG2a
  • 轻链类型

    kappa
  • 研究领域

    • 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

相关产品

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG2a, kappa monoclonal [MG2a-53] - Isotype control (ab18415)
  • KO cell lines

    • Human SQSTM1 (p62) knockout HEK-293T cell line (ab255343)
  • KO cell lysates

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

    • Human SQSTM1 (p62) knockout HEK293T cell pellet (ab278927)
  • Recombinant Protein

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

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

应用

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab56416 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用 Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
WB
Human
应用 Ab评论 说明
IHC-P (3)
Use at an assay dependent concentration.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

WB (22)
Use at an assay dependent concentration.
ICC/IF (4)
Use at an assay dependent concentration.

Fix cells for 15 minutes with 2 mL of 4% paraformaldehyde solution (pH 7.4 with NaOH in PBS). Permeabilize cells by incubating for 15 minutes on ice with 2 mL of 0.1% Triton X-100 in PBS.

Flow Cyt
Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

We recommend Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) secondary antibody

说明
IHC-P
Use at an assay dependent concentration.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

WB
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.

Fix cells for 15 minutes with 2 mL of 4% paraformaldehyde solution (pH 7.4 with NaOH in PBS). Permeabilize cells by incubating for 15 minutes on ice with 2 mL of 0.1% Triton X-100 in PBS.

Flow Cyt
Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

We recommend Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) secondary antibody

靶标

  • 功能

    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.
  • 组织特异性

    Ubiquitously expressed.
  • 疾病相关

    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.
  • 序列相似性

    Contains 1 OPR domain.
    Contains 1 UBA domain.
    Contains 1 ZZ-type zinc finger.
  • 结构域

    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.
  • 翻译后修饰

    Phosphorylated. May be phosphorylated by PRKCZ (By similarity). Phosphorylated in vitro by TTN.
  • 细胞定位

    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
  • 数据库链接

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

      • 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

    图片

    • Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)
      Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)
      All lanes : Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416) at 1/1000 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : SQSTM1 knockout HEK293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

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



      Lanes 1- 2: Merged signal (red and green). Green - ab56416 observed at 62 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

       ab56416 was shown to react with SQSTM1/p62 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 HEK293T and SQSTM1 knockout HEK293T cell lysates 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) overnight at 4°C at a 1 in 1000 dilution and a 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)
      Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)

      Monoclonal antibody to SQSTM1 (ab56416) on HeLa cell, antibody concentration 10 ug/ml.

      This image was generated using the ascites version of the product.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)

      ab56416 (1µg/ml) staining SQSTM1 in human lymph node using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear and cytoplasmic staining.
      Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

      This image was generated using the ascites version of the product.

    • Flow Cytometry - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)
      Flow Cytometry - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab56416)

      Overlay histogram showing HeLa cells stained with ab56416 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56416, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

      This image was generated using the ascites version of the product.

    实验方案

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

    Click here to view the general protocols

    数据表及文件

    • Datasheet
    • SDS
  • 文献 (499)

    发表研究结果有使用 ab56416?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab56416 被引用在 499 文献中.

    • Zhang J  et al. Combination therapy with ropivacaine-loaded liposomes and nutrient deprivation for simultaneous cancer therapy and cancer pain relief. Theranostics 10:4885-4899 (2020). PubMed: 32308756
    • Zhang L  et al. Ethionine Suppresses Mitochondria Autophagy and Induces Apoptosis via Activation of Reactive Oxygen Species in Neural Tube Defects. Front Neurol 11:242 (2020). PubMed: 32318018
    • Zhu N  et al. Berberine Protects Against Simulated Ischemia/Reperfusion Injury-Induced H9C2 Cardiomyocytes Apoptosis In Vitro and Myocardial Ischemia/Reperfusion-Induced Apoptosis In Vivo by Regulating the Mitophagy-Mediated HIF-1a/BNIP3 Pathway. Front Pharmacol 11:367 (2020). PubMed: 32292345
    • Chen X  et al. miR-155 facilitates calcium oxalate crystal-induced HK-2 cell injury via targeting PI3K associated autophagy. Exp Mol Pathol 115:104450 (2020). PubMed: 32417393
    • Buch BT  et al. Colchicine treatment impairs skeletal muscle mitochondrial function and insulin sensitivity in an age-specific manner. FASEB J 34:8653-8670 (2020). PubMed: 32372536
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价

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    1-10 of 35 Abreviews

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-SQSTM1 / p62 antibody - Autophagosome Marker

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (hCRC)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: tris pH9
    Permeabilization
    Yes - Tween-20 (1%)
    Specification
    hCRC
    Blocking step
    BSA/FBS in TBS as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: rt°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Jul 14 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-SQSTM1 / p62 antibody - Autophagosome Marker

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Mouse breast cancer)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: tris pH9
    Permeabilization
    Yes - Tween-20 (1%)
    Specification
    Mouse breast cancer
    Blocking step
    No blocking step used for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: rt°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Jul 14 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-SQSTM1 / p62 antibody - Autophagosome Marker

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (CAFs)
    Permeabilization
    Yes - Tween-20 (1%)
    Specification
    CAFs
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Jul 07 2020

    Western blot abreview for Anti-SQSTM1 / p62 antibody - Autophagosome Marker

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (CAFs)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    50 µg
    Specification
    CAFs
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jul 06 2020

    Western blot abreview for Anti-SQSTM1 / p62 antibody - Autophagosome Marker

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Hematological cancer cell line lysates)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    30 µg
    Specification
    Hematological cancer cell line lysates
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jan 14 2020

    IHC - Wholemount abreview for Anti-SQSTM1 / p62 antibody - Autophagosome Marker

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    IHC - Wholemount
    Sample
    Mouse Tissue (Retinal pigment epithelium/chroid complex)
    Specification
    Retinal pigment epithelium/chroid complex
    Read More

    Dr. Jingyu Yao

    Verified customer

    提交于 Jan 17 2019

    Western blot abreview for Anti-SQSTM1 / p62 antibody - Autophagosome Marker

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (neuroblastoma)
    Gel Running Conditions
    Reduced Denaturing (13.5)
    Loading amount
    20 µg
    Specification
    neuroblastoma
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    提交于 Dec 14 2018

    Immunocytochemistry abreview for Anti-SQSTM1 / p62 antibody - Autophagosome Marker

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Rat Cultured Cells (H9C2 rat cardiomyocytes)
    Permeabilization
    Yes - Triton x-100, 0.03%
    Specification
    H9C2 rat cardiomyocytes
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Nov 13 2018

    Western blot abreview for Anti-SQSTM1 / p62 antibody - Autophagosome Marker

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (DU145 Prostate cancer cell line)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    DU145 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

    Dr. Dimitra Kalamida

    Verified customer

    提交于 Jul 30 2018

    Western blot abreview for Anti-SQSTM1 / p62 antibody - Autophagosome Marker

    Excellent
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Rat Cell lysate - other (neuron)
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Loading amount
    15 µg
    Treatment
    different concentration of poisoning for 24hrs
    Specification
    neuron
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jan 24 2018

    1-10 of 35 Abreviews

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    研究类别(A-Z)
    • 肿瘤研究
    • 心血管研究
    • 细胞生物学
    • 发育生物学
    • 染色质及细胞核信号转导研究
    • 免疫学
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