P62,SQSTM1 Recombinant antibody

• Featured Product
• KD/KO Validated

P62,SQSTM1 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

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

Conjugate

Unconjugated

Publications

3

CloneNo.

1D17

Cat no : 80294-1-RR

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 80294-1-RR

  Lysates of HepG2 cells treated with Chloroquine or not were subjected to SDS PAGE followed by western blot with 80294-1-RR (P62,SQSTM1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 80294-1-RR

  WB result of P62,SQSTM1 antibody (80294-1-RR; 1:600; 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 using 80294-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 80294-1-RR (P62,SQSTM1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of HepG2 using 80294-1-RR

  WB result of P62,SQSTM1 antibody (80294-1-RR; 1:600; incubated at room temperature for 1.5 hours) with sh-Control and sh-P62,SQSTM1 transfected HepG2 cells.

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  IP experiment of HEK-293 using 80294-1-RR

  IP result of anti-P62,SQSTM1 (IP:80294-1-RR, 4ug; Detection:80294-1-RR 1:500) with HEK-293 cells lysate 1640 ug.

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  IHC staining of human pancreas cancer using 80294-1-RR

  Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue slide using 80294-1-RR (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 stomach cancer using 80294-1-RR

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 80294-1-RR (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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  IF Staining of HeLa using 80294-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed Chloroquine treated HeLa cells using P62,SQSTM1 antibody (80294-1-RR, Clone: 1D17 ) at dilution of 1:500 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

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  FC experiment of Jurkat using 80294-1-RR

  1X10^6 Jurkat cells were intracellularly stained with 0.4 ug Anti-Human P62,SQSTM1 (80294-1-RR, Clone:1D17) and CoraLite®488-Conjugated 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: HepG2 cells, HEK-293 cells, MCF-7 cells, HeLa cells
• Positive IP detected in: HEK-293 cells
• Positive IHC detected in: human pancreas cancer tissue, human stomach 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 HeLa cells
• Positive FC (Intra) detected in: Jurkat cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:250-1:1000
• 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

• WB: See 3 publications below

Product Information

80294-1-RR targets P62,SQSTM1 in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• 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_2918883
• 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 aggregated proteins for autophagic degradation. p62 is degraded during the autophagic process, which makes its intracellular level a marker for autophagy progression. 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, and nephropathic cystinosis (PMID: 22074114, 22860231, 22714671).

Publications

Species	Application	Title
human	WB	Toxicol In Vitro; Paraquat-induced ferroptosis suppression via NRF2 expression regulation; Authors - Qiqi Cai
human	WB	Front Pharmacol; Inhibition of NNMT enhances drug sensitivity in lung cancer cells through mediation of autophagy; Authors - Jian Wang
human	WB	Oncogene; Suppression of lysosome metabolism-meditated GARP/TGF-β1 complexes specifically depletes regulatory T cells to inhibit breast cancer metastasis; Authors - Jing Ma