Anticorps Monoclonal anti-P62,SQSTM1

• Phare
• Validé par KD/KO

P62,SQSTM1 Monoclonal Antibody for FC, IF, IHC, IP, WB, ELISA

Hôte / Isotype

Mouse / IgG2b

Réactivité testée

Humain et plus (3)

Applications

WB, IP, IHC, IF, FC, CoIP, ELISA

Conjugaison

Non conjugué

Publications

104

CloneNo.

1H5C1

Avis (1)

N° de cat : 66184-1-Ig

Synonymes

A170, EBIAP, ORCA, OSIL, p60, P62, P62/SQSTM1, P62;SQSTM1, p62B, PDB3, sequestosome 1, SQSTM1, Ubiquitin binding protein p62, ZIP3

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Galerie de données de validation

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  WB analysis using 66184-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66184-1-Ig (P62,SQSTM1 antibody) at dilution of 1:40000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

• 

  WB analysis of HEK-293 using 66184-1-Ig

  WB result of P62,SQSTM1 antibody (66184-1-Ig; 1:1000; incubated at room temperature for 1.5 hours) with sh-Control and sh-P62/SQSTM1 transfected HEK-293 cells.

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  WB analysis of U2OS using 66184-1-Ig

  U2OS (WT and SQSTM1 KO) lysates prepared with RIPA buffer, 25 μg protein loaded. 66184-1-Ig incubated at 1:1000 at 4°C overnight in 5% milk in TBST. Ponceau stained transfers shown on right. Data provided by YCharOS, an open science company with a mission to validate commercial antibodies to improve scientific reproducibility and transparency.

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  WB analysis of K-562 using 66184-1-Ig

  K-562 cells were subjected to SDS PAGE followed by western blot with 66184-1-Ig (P62/SQSTM1 antibody) at dilution of 1:40000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of U2OS using 66184-1-Ig

  U2OS lysates prepared and IP of SQSTM1 performed using 1.0 μg of 66184-1-Ig coupled to protein G- Sepharose beads. The Ponceau stained transfers of each blot are shown. Data provided by YCharOS, an open science company with a mission to validate commercial antibodies to improve scientific reproducibility and transparency.

• 

  IHC staining of human lung cancer using 66184-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66184-1-Ig (P62,SQSTM1 antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver cancer using 66184-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66184-1-Ig (P62,SQSTM1 antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human colon cancer using 66184-1-Ig

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 66184-1-Ig (P62,SQSTM1 antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human endometrial cancer using 66184-1-Ig

  Immunohistochemical analysis of paraffin-embedded human endometrial cancer tissue slide using 66184-1-Ig (P62,SQSTM1 antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver cancer using 66184-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66184-1-Ig (P62/SQSTM1 antibody at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of U2OS using 66184-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed U2OS cells using P62/SQSTM1 antibody (66184-1-Ig, Clone: 1H5C1 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), CL594-Phalloidin (red).

• 

  IF Staining of U2OS using 66184-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed Chloroquine treated U2OS cells using P62/SQSTM1 antibody (66184-1-Ig, Clone: 1H5C1 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), CL594-Phalloidin (red).

• 

  IF Staining of HepG2 using 66184-1-Ig

  Immunofluorescent analysis of (-20℃ Ethanol ) fixed Starvation treated HepG2 cells using 66184-1-Ig(P62,SQSTM1 antibody) at dilution of 1:100 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• 

  FC experiment of Jurkat using 66184-1-Ig

  1X10^6 Jurkat cells were stained with 0.20ug P62,SQSTM1 antibody (66184-1-Ig, red) and control antibody (blue). Fixed with 90% MeOH.

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Applications testées

• Résultats positifs en WB: cellules HeLa, cellules HEK-293, cellules HepG2, cellules Jurkat, cellules K-562, cellules L02, cellules MCF-7, cellules Raji, cellules U2OS
• Résultats positifs en IP: cellules U2OS,
• Résultats positifs en IHC: tissu de cancer du poumon humain, tissu de cancer de l'endomètre humain, tissu de cancer du côlon humain, tissu de cancer du foie humain; il est suggéré de démasquer l'antigène avec un tampon de TE buffer pH 9.0; (*) À défaut, 'le démasquage de l'antigène peut être 'effectué avec un tampon citrate pH 6,0.
• Résultats positifs en IF: cellules U2OS, cellules HepG2 traitées par déprivation, cellules U2OS traitées à la chloroquine
• Résultats positifs en cytométrie: cellules Jurkat,

Dilution recommandée

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprécipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochimie (IHC): IHC : 1:2000-1:8000
• Immunofluorescence (IF): IF : 1:200-1:800
• Flow Cytometry (FC): FC : 1:10-1:100

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, check data in validation data gallery

Applications publiées

• WB: See 96 publications below
• IHC: See 9 publications below
• IF: See 15 publications below
• IP: See 2 publications below
• CoIP: See 2 publications below

Informations sur le produit

66184-1-Ig cible P62,SQSTM1 dans les applications de WB, IP, IHC, IF, FC, CoIP, ELISA et montre une réactivité avec des échantillons Humain

• Réactivité: Humain
• Réactivité citée: bovin, Humain, porc, singe
• Hôte / Isotype: Mouse / IgG2b
• Clonalité: Monoclonal
• Type: Anticorps
• Immunogène: P62,SQSTM1 Protéine recombinante Ag13131
• Nom complet: sequestosome 1
• Masse moléculaire calculée: 48 kDa
• Poids moléculaire observé: 62 kDa
• Numéro d’acquisition GenBank: BC017222
• Symbole du gène: SQSTM1
• Identification du gène (NCBI): 8878
• Conjugaison: Non conjugué
• Forme: Liquide
• Méthode de purification: Purification par protéine A
• Tampon de stockage: PBS avec azoture de sodium à 0,02 % et glycérol à 50 % pH 7,3
• Conditions de stockage: Stocker à -20°C. Stable pendant un an après l'expédition. L'aliquotage n'est pas nécessaire pour le stockage à -20^oC Les 20ul contiennent 0,1% de BSA.

Informations générales

Sequestosome 1 (SQSTM1/p62) is a multifunctional adaptor protein implicated in selective autophagy, cell signaling pathways, and tumorigenesis. p62 has been implicated in shuttling ubiquitinated and sometimes aggregated proteins for autophagic degradation. As a autophagy-specific substrate, p62 is degraded during the autophagic process, which makes intracellular level of p62 as a marker for autophagy flux. p62 is at the cross-roads of several signaling pathways including Ras/ Raf/ MAPK and NFκB and plays important role in cancer. p62 is a component of inclusion bodies/ protein aggregates found in human diseases, including Huntington's disease, Alzheimer's disease, Parkinson's disease in the brain, and nephropathic cystinosis in kidney (PMID: 22074114, 22860231, 22714671). The molecular weight of p62 is predicted as 48/ 38 kDa, while western blot analyses using this antibody demonstrate the major band around 60-62 kDa in various tissues.

Publications

Species	Application	Title
human	WB,IHC	Gastroenterology; Proteomic characterization identifies clinically relevant subgroups of gastrointestinal stromal tumors; Authors - Mingjun Sun
human	IHC,IF	Acta Neuropathol; Upstream open reading frame with NOTCH2NLC GGC expansion generates polyglycine aggregates and disrupts nucleocytoplasmic transport: implications for polyglycine diseases.; Authors - Shaoping Zhong
human	WB	Nat Commun; IL-6 regulates autophagy and chemotherapy resistance by promoting BECN1 phosphorylation.; Authors - Fuqing Hu
	IHC	Mol Neurodegener; Collusion of α-Synuclein and Aβ aggravating co-morbidities in a novel prion-type mouse model; Authors - Grace M. Lloyd
human	WB	J Cell Biol; CHMP2B regulates TDP-43 phosphorylation and cytotoxicity independent of autophagy via CK1.; Authors - Xue Deng
human	WB	Theranostics; The c-Myc/miR-27b-3p/ATG10 regulatory axis regulates chemoresistance in colorectal cancer.; Authors - Wu Sun

Avis

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.

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FH

David (Verified Customer) (01-13-2020)

Used for immunocytochemistry. Low signal in control cells, but very strong response when treated with bafilomycin A. Very obvious p62 puncta in cells.

• Applications: Immunofluorescence,
• Primary Antibody Dilution: 1:100
• Cell Tissue Type: SH-SY5Y

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