P62,SQSTM1 Polyclonal antibody

• Featured Product
• KD/KO Validated

P62,SQSTM1 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human and More (7)

Applications

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

Conjugate

Unconjugated

Publications

1633

Reviews (7)

Cat no : 18420-1-AP

Synonyms

P62/SQSTM1, P62, SQSTM1, Sequestosome-1, Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa

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Validation Data Gallery

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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.

• 

  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.

• 

  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.

• 

  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.

• 

  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.

• 

  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.

• 

  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).

• 

  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).

• 

  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).

• 

  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).

• 

  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).

• 

  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).

• 

  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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Tested Applications

• Positive WB detected in: HeLa cells, U2OS cells, HEK-293 cells, HepG2 cells, U-87 MG cells, Jurkat cells
• Positive IP detected in: HepG2 cells, U2OS cells
• Positive IHC detected in: human lung cancer tissue, human gliomas tissue, human liver cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: Chloroquine treated HepG2 cells, U2OS cells, Chloroquine treated HeLa cells
• Positive FC (Intra) detected in: HEK-293 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:200-1:800
• Immunofluorescence (IF)/ICC: IF/ICC : 1:750-1:3000
• Flow Cytometry (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.

Published Applications

• KD/KO: See 36 publications below
• WB: See 1501 publications below
• IHC: See 154 publications below
• IF: See 183 publications below
• IP: See 22 publications below
• CoIP: See 9 publications below

Product Information

18420-1-AP targets P62,SQSTM1 in WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human, rabbit, monkey, chicken, zebrafish, hamster, goat, goslings
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: P62,SQSTM1 fusion protein Ag13131
• Full Name: sequestosome 1
• Calculated Molecular Weight: 48 kDa
• Observed Molecular Weight: 62 kDa
• GenBank Accession Number: BC017222
• Gene Symbol: P62/SQSTM1
• Gene ID (NCBI): 8878
• RRID: AB_10694431
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

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. 

Publications

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	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
human	WB	Protein Cell; ORP8 acts as a lipophagy receptor to mediate lipid droplet turnover; Authors - Maomao Pu
	WB	Cell Res; NudCL2 is an autophagy receptor that mediates selective autophagic degradation of CP110 at mother centrioles to promote ciliogenesis.; Authors - Min Liu

Reviews

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)

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