概述

  • 产品名称

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

    小鼠单克隆抗体to SQSTM1 / p62
  • 宿主

    Mouse
  • 经测试应用

    适用于: IHC-P, WB, ICC/IF, Flow Cyt, IHC-Frmore details
  • 种属反应性

    与反应: Mouse, Rat, Human, Rhesus monkey, Chinese hamster
  • 免疫原

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

  • 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.

    Abcam recommended secondaries - Goat Anti-Mouse HRP (ab205719) and Goat Anti-Mouse Alexa Fluor® 488 (ab150113).

    See other anti-mouse secondary antibodies that can be used with this antibody.

性能

应用

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.

应用 Ab评论 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

IHC-Fr Use at an assay dependent concentration. PubMed: 22577215

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.
  • Information by UniProt
  • 数据库链接

  • 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

  • Lane 1: Hap1 wildtype cell lysate (20 µg)

    Lane 2: SQSTM1 Hap1 knockout cell lysate (20 µg)

    Lane 3: HeLa wildtype cell lysate (20 µg)

    Lane 4: SQSTM1 HeLa knockout cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab56416 observed at 62 kDa. Red - loading control, ab181602 observed at 37 kDa.

    ab56416 was shown to react with SQSTM1 / p62 in HeLa wildtype. Loss of signal was observed when knockout sample ab263770 was used. Wild-type and SQSTM1 / p62 knockout samples were subjected to SDS-PAGE. 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-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.
  • 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.

  • All lanes : Anti-SQSTM1 / p62 antibody (ab56416) at 1/2000 dilution

    All lanes : HEK293 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-mouse IgG at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 61 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocked with 5% milk for 3 hours at 21°C.

    Incubated with the primary antibody for 17 hours at 4°C.

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

    See Abreview

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

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

  • ab56416 staining SQSTM1/p62 in Human A431 epidermoid cancer cells by ICC/IF (Immunocytochemistry/immunoflurescence). Cells were fixed with formaldehyde, permeabilized with Triton X-100 and blocked with 5% BSA for 30 minutes at room temperature.  Samples were incubated with  primary antibody (1/50) in 5% BSA for 1 hour. An Alexa Fluor® 488-conjugated Goat monoclonal to mouse IgG (1/50) was used as secondary antibody. 

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

    See Abreview

  • 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.

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

    Lane 1 : Control
    Lane 2 : Starved in HBSS for 2 hours
    Lane 3 : Starved in HBSS for 4 hours
    Lane 4 : Starved in HBSS for 8 hours
    Lane 5 : Starved in HBSS 8 hours + 200 nM Baf A1
    Lane 6 : Starved in HBSS 8 hours + 4 uM Mg132

    Lysates/proteins at 40 µg per lane.

    Secondary
    All lanes : HRP conjugated goat anti-mouse polyclonal at 1/5000 dilution

    Developed using the ECL technique.

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


    Exposure time: 10 seconds


    All lanes are whole cell lysate prepared from HeLa cells. Treatments are listed.

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

    See Abreview

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

    Lane 1 : Whole cell lysates prepared from Tzb-naive SKBR3 parental cells.
    Lane 2 : Whole cell lysates prepared from Tzb-refractory TzbR POOL1 cells.
    Lane 3 : Whole cell lysates prepared from Tzb-refractory TzbR POOL2 cells.

    Lysates/proteins at 50 µg per lane.

    Secondary
    All lanes : Horseradish peroxidase-conjugated secondary

    Developed using the ECL technique.


    Cells were washed twice with cold-PBS and then lysed in buffer (20 mM Tris pH 7.5, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% Triton® X-100, 2.5 mM sodium pyrophosphate, 1 mM ß-glycerolphosphate, 1 mM Na3VO4, 1 µg/mL leupeptin, 1 mM phenylmethylsulfonylfluoride, and complete protease inhibitor cocktail for 30 minutes on ice. The lysates were cleared by centrifugation in an Eppendorff tube (15 minutes at 14,000×g, 4°C). Protein content was determined against a standardized control using the Pierce Protein Assay Kit. Equal amounts of protein were resuspended in 5× Laemmli sample buffer (10 minutes at 70°C), resolved by electrophoresis on 10% SDS-PAGE, and transferred onto nitrocellulose membranes. Non-specific binding on the nitrocellulose filter paper was minimized by blocking for 1 hour at room temperature with TBS-T buffer [25 mM Tris-HCl (pH 7.5), 150 mM NaCl, 0.05% Tween 20] containing 5% (w/v) nonfat dry milk. The treated filters were washed in TBS-T and then incubated with the primary

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

文献

This product has been referenced in:

  • King BC  et al. Complement Component C3 Is Highly Expressed in Human Pancreatic Islets and Prevents ß Cell Death via ATG16L1 Interaction and Autophagy Regulation. Cell Metab 29:202-210.e6 (2019). Read more (PubMed: 30293775) »
  • Yamasaki TR  et al. Parkinson's disease and multiple system atrophy have distinct a-synuclein seed characteristics. J Biol Chem 294:1045-1058 (2019). Read more (PubMed: 30478174) »
See all 358 Publications for this product

客户评价及客户问答

1-10 of 41 Abreviews or Q&A

Application
IHC - Wholemount
Sample
Mouse Tissue (Retinal pigment epithelium/chroid complex)
Specification
Retinal pigment epithelium/chroid complex

Dr. Jingyu Yao

Verified customer

提交于 Jan 17 2019

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

Abcam user community

Verified customer

提交于 Dec 14 2018

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

Abcam user community

Verified customer

提交于 Nov 13 2018

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

Dr. Dimitra Kalamida

Verified customer

提交于 Jul 30 2018

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

Abcam user community

Verified customer

提交于 Jan 24 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (liver, HepG2)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Specification
liver, HepG2
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

提交于 Dec 04 2017

Application
Western blot
Sample
Monkey Tissue lysate - whole (cardiac)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
cardiac
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

提交于 Nov 01 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293 cells)
Gel Running Conditions
Reduced Denaturing (12,5 %)
Loading amount
20 µg
Specification
HEK293 cells
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Dr. Melanie Thelen

Verified customer

提交于 May 23 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (brain, liver, RPE)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Treatment
5 ug 4-HNE per eye
Specification
brain, liver, RPE
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

提交于 Mar 22 2017

Application
Western blot
Sample
Mouse Cell lysate - whole cell (in vitro differentiated neuronal cells)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
20 µg
Treatment
100 ng/ml chloroquine for 4 hr
Specification
in vitro differentiated neuronal cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

提交于 Mar 10 2017

1-10 of 41 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

注册