P62,SQSTM1 Monoclonal antibody

• Featured Product
• KD/KO Validated

P62,SQSTM1 Monoclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

human and More (3)

Applications

WB, IHC, IF/ICC, IP, CoIP, ELISA

Conjugate

Unconjugated

Publications

145

CloneNo.

1H5C1

Reviews (2)

Cat no : 66184-1-Ig

Synonyms

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

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Validation Data Gallery

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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 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.

• 

  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.

• 

  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 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).

• 

  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).

• 

  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 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 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 Goat Anti-Mouse IgG(H+L).

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Tested Applications

• Positive WB detected in: HeLa cells, U2OS cells, K-562 cells, HEK-293 cells, MCF-7 cells, HepG2 cells, Jurkat cells, L02 cells, Raji cells
• Positive IP detected in: U2OS cells
• Positive IHC detected in: human lung cancer tissue, human colon cancer tissue, human liver cancer tissue, human endometrial 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: U2OS cells, Chloroquine treated U2OS cells, Starvation treated HepG2 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 : 1:2000-1:8000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800

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

• WB: See 132 publications below
• IHC: See 10 publications below
• IF: See 24 publications below
• IP: See 3 publications below
• CoIP: See 4 publications below

Product Information

66184-1-Ig targets P62,SQSTM1 in WB, IHC, IF/ICC, IP, CoIP, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human, pig, monkey, bovine
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• 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_2881579
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A 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

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

Reviews

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

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FH

Eleonora (Verified Customer) (04-12-2024)

The antibody unfortunately did not work in IF, no signal. In western blot the band is present but very faint.

• Applications: Western Blot, Immunofluorescence
• Primary Antibody Dilution: western blot 1:500. IF 1:200
• Cell Tissue Type: iPSC-derived neurons

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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