P62/SQSTM1 Polyklonaler Antikörper

• Featured Product
• KD/KO Validated

P62/SQSTM1 Polyklonal Antikörper für WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Wirt / Isotyp

Kaninchen / IgG

Getestete Reaktivität

human und mehr (6)

Anwendung

WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA

Konjugation

Unkonjugiert

Publikationen(1897)

Rezensionen (7)

Kat-Nr. : 18420-1-AP

Synonyme

P62,SQSTM1, P62, SQSTM1, EBI3-associated protein of 60 kDa, Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa

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  WB analysis using 18420-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 18420-1-AP (P62,SQSTM1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis using 18420-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 18420-1-AP (P62,SQSTM1 antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours.

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  WB analysis of U2OS using 18420-1-AP

  U2OS (WT and SQSTM1 KO) lysates prepared with RIPA buffer, 25 μg protein loaded. 18420-1-AP 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 HEK-293 using 18420-1-AP

  WB result of P62,SQSTM1 antibody (18420-1-AP; 1:20000; incubated at room temperature for 1.5 hours) with sh-Control and sh-P62/SQSTM1 transfected HEK-293 cells.

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  IP experiment of U2OS using 18420-1-AP

  U2OS lysates prepared and IP of SQSTM1 performed using 1.0 μg of 18420-1-AP coupled to protein A- 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.

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  IP experiment of HepG2 using 18420-1-AP

  IP result of anti-P62,SQSTM1 (IP:18420-1-AP, 4ug; Detection:18420-1-AP 1:6000) with HepG2 cells lysate 1360 ug.

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  IHC staining of human gliomas using 18420-1-AP

  Immunohistochemical analysis of paraffin-embedded human gliomas using 18420-1-AP (SQSTM1 antibody) at dilution of 1:50 (under 40x lens).

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  IHC staining of human liver cancer using 18420-1-AP

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 18420-1-AP (P62,SQSTM1 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver cancer using 18420-1-AP

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 18420-1-AP (P62,SQSTM1 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lung cancer using 18420-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 18420-1-AP (P62,SQSTM1 antibody) at dilution of 1:400 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 18420-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 18420-1-AP (P62,SQSTM1 antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 18420-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 18420-1-AP (P62,SQSTM1 antibody) at dilution of 1:400 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 18420-1-AP

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 18420-1-AP (P62,SQSTM1 antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver cancer using 18420-1-AP

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 18420-1-AP (P62/SQSTM1 antibody at dilution of 1:50 (under 10x lens).

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  IF Staining of HepG2 using 18420-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed Chloroquine treated HepG2 cells using P62,SQSTM1 antibody (18420-1-AP) at dilution of 1:1500 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

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  IF Staining of HeLa using 18420-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed Chloroquine treated HeLa cells using P62,SQSTM1 antibody (18420-1-AP) at dilution of 1:500 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), (CL594-Phalloidin, red).

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  IF Staining of U2OS using 18420-1-AP

  U2OS WT cells (yellow outline) and SQSTM1 KO cells (red outline) labelled with a green or a far-red fluorescence dye, respectively. Cells fixed with 4% PFA and stained with 18420-1-AP at 1:300. Bars = 10 μm. Data provided by YCharOS, an open science company with a mission to validate commercial antibodies to improve scientific reproducibility and transparency.

• 

  FC experiment of HEK-293 using 18420-1-AP

  1X10^6 HEK-293 cells were intracellularly stained with 0.4 ug Anti-Human P62,SQSTM1 (18420-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Isotype Control. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Geprüfte Anwendungen

• Erfolgreiche Detektion in WB: HeLa-Zellen, HEK-293-Zellen, HepG2-Zellen, Jurkat-Zellen, U2OS-Zellen, U-87 MG-Zellen
• Erfolgreiche IP: HepG2-Zellen, U2OS-Zellen
• Erfolgreiche Detektion in IHC: humanes Lungenkarzinomgewebe, humanes Gliomgewebe, humanes Leberkarzinomgewebe; Hinweis: Antigendemaskierung mit TE-Puffer pH 9,0 empfohlen. (*) Wahlweise kann die Antigendemaskierung auch mit Citratpuffer pH 6,0 erfolgen.
• Erfolgreiche Detektion in IF/ICC: mit Chloroquin behandelte HepG2-Zellen, mit Chloroquin behandelte HeLa-Zellen, U2OS-Zellen
• Erfolgreiche Detektion in FC (Intra): HEK-293-Zellen

Empfohlene Verdünnung

• Western Blot (WB): WB : 1:5000-1:50000
• Immunpräzipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunhistochemie (IHC): IHC : 1:200-1:800
• Immunfluoreszenz (IF)/ICC: IF/ICC : 1:750-1:3000
• Durchflusszytometrie (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

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

Veröffentlichte Anwendungen

• KD/KO: See 41 publications below
• WB: See 1731 publications below
• IHC: See 168 publications below
• IF: See 218 publications below
• IP: See 23 publications below
• CoIP: See 13 publications below

Produktinformation

18420-1-AP bindet in WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA P62/SQSTM1 und zeigt Reaktivität mit human

• Getestete Reaktivität: human
• In Publikationen genannte Reaktivität: human, Affe, Gänseküken, Huhn, Kaninchen, Zebrafisch, Ziege
• Wirt / Isotyp: Kaninchen / IgG
• Klonalität: Polyklonal
• Typ: Antikörper
• Immunogen: P62/SQSTM1 fusion protein Ag13131
• Vollständiger Name: sequestosome 1
• Berechnetes Molekulargewicht: 48 kDa
• Beobachtetes Molekulargewicht: 62 kDa
• GenBank-Zugangsnummer: BC017222
• Gene symbol: P62/SQSTM1
• Gene ID (NCBI): 8878
• Konjugation: Unkonjugiert
• Form: Liquid
• Reinigungsmethode: Antigen-Affinitätsreinigung
• Lagerungspuffer: PBS with 0.02% sodium azide and 50% glycerol
• Lagerungsbedingungen: Bei -20°C lagern. Nach dem Versand ein Jahr lang stabil Aliquotieren ist bei -20^oC Lagerung nicht notwendig. 20ul Größen enthalten 0,1% BSA.

Hintergrundinformationen

Background

 P62 (ubiquitin-binding protein P62), also known as Sequestosome-1, is a multifunctional adaptor protein most widely known for its role as an autophagosome cargo protein (PMID: 8551575). P62 via specific interactions with polyubiquitylated target proteins induces their selective autophagy (PMID: 17580304). It also plays an important role in the regulation of the NFkB signaling pathway, senescence, cell differentiation, apoptosis, and immune responses (PMID: 26404812).

 What is the molecular weight of P62?

 The observed molecular weight of the protein can vary from as low as 8 kDa (for the smallest isoforms) to 48 kDa.

 What is the subcellular localization of P62?

 P62 is mainly localized in the cytoplasm; however, upon autophagy induction, e.g., via starvation or selective inhibitor treatment, it localizes in vesicular structures - autophagosomes.

 What is the tissue specificity of P62?

 It is ubiquitously expressed in various tissues.

 What is the function of P62 in the regulation of cell death and autophagy?

 It is a selective autophagy receptor that forms a bridge between polyubiquitylated cargo (via its UBA domains) and an autophagy modifier such as LC3 (via LIR domains) (PMIDs: 16286508, 20168092, 24128730, 28404643, 22622177). The process of selective autophagy is tightly regulated at many levels, including the posttranslational modifications (PTMs) of various proteins in the cascade, P62 among others (PMID: 29233872). P62 is involved in the regulation of cell death induction in response to various stimuli, e.g., via activation of caspase-8 at the autophagosome membrane (PMID: 29480462). In addition, P62 is degraded during the autophagic process, which makes its intracellular level a marker for autophagy progression.

 What is P62's involvement in disease?

 Mutations in P62 have been associated with the following diseases: sporadic and familial Paget's disease of bone, neurodegenerative diseases, diabetes, and obesity (PMID: 29480462). A growing number of reports suggest the implication of P62 in the induction of multiple cellular oncogenic transformations. Indeed, increased levels of P62 have been linked to tumor formation, cancer promotion, and resistance to therapy (PMID: 29738493). Moreover, P62 is an unfavorable prognostic marker in liver cancer. 

Publikationen

Species	Application	Title
human	WB	Nature; FBXO38 mediates PD-1 ubiquitination and regulates anti-tumour immunity of T cells.; Authors - Xiangbo Meng
human	WB	Signal Transduct Target Ther; Targeting CRL4 suppresses chemoresistant ovarian cancer growth by inducing mitophagy; Authors - Yang Meng
human	WB	Mol Cancer; Overcoming multi-drug resistance in SCLC: a synergistic approach with venetoclax and hydroxychloroquine targeting the lncRNA LYPLAL1-DT/BCL2/BECN1 pathway; Authors - Shuxin Li
	WB	Cell Metab; Disrupted methionine cycle triggers muscle atrophy in cancer cachexia through epigenetic regulation of REDD1; Authors - Kai Lin
human	WB	Cancer Cell; iASPP Is an Antioxidative Factor and Drives Cancer Growth and Drug Resistance by Competing with Nrf2 for Keap1 Binding.; Authors - Wenjie Ge
	WB	Cell Metab; Receptor-Mediated ER Export of Lipoproteins Controls Lipid Homeostasis in Mice and Humans.; Authors - Xiao Wang

Rezensionen

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

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FH

David (Verified Customer) (01-02-2024)

Used for WB in cell lysates. Prominent band at the predicted molecular weight, although some non-specific bands in the control at around 40kDa. However, the correct SQSTM1 band can be clearly visualised.

• Applications: Western Blot
• Primary Antibody Dilution: 1:500
• Cell Tissue Type: SH-SY5Y lysate

FH

Alexandru (Verified Customer) (11-21-2023)

Great at detecting p62 in cell lysates.

• Applications: Western Blot
• Cell Tissue Type: HEK-293

FH

Wan (Verified Customer) (02-24-2022)

Perfect antibody, clear bands!

• Applications: Western Blot

FH

X (Verified Customer) (01-18-2021)

It is a reliable Ab to detect p62 with mouse brain lysate, although there is other band detected.

• Applications: Western Blot
• Primary Antibody Dilution: 1:2000
• Cell Tissue Type: mouse brain

P62/SQSTM1 Antibody Western Blot validation (1:2000 dilution) in mouse brain (Cat no:18420-1-AP)

FH

X (Verified Customer) (01-18-2021)

It is a reliable Ab to detect p62 with mouse brain lysate in WB, although there is other bands seen.

• Applications: Western Blot
• Primary Antibody Dilution: 1:2000
• Cell Tissue Type: mouse brain

FH

Wei (Verified Customer) (01-30-2020)

Good, Clear band for mouse heart

• Applications: Western Blot,
• Cell Tissue Type: Mouse Heart

FH

Joleen (Verified Customer) (06-16-2019)

Recognizes SQSTM1 prominently with some non-specific bands.

• Applications: Western Blot,
• Primary Antibody Dilution: 1:1k
• Cell Tissue Type: MDCK GII

P62/SQSTM1 Antibody Western Blot, validation (1:1k dilution) in MDCK GII (Cat no:18420-1-AP)