P62/SQSTM1 Monoclonal antibody, PBS Only

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  WB analysis using 66184-1-Ig (same clone as 66184-1-PBS)

  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. This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

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  WB analysis of U2OS using 66184-1-Ig (same clone as 66184-1-PBS)

  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. This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

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  WB analysis of HEK-293 using 66184-1-Ig (same clone as 66184-1-PBS)

  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. This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

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  WB analysis of K-562 using 66184-1-Ig (same clone as 66184-1-PBS)

  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. This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

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  IP experiment of U2OS using 66184-1-Ig (same clone as 66184-1-PBS)

  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. This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

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  IHC staining of human lung cancer using 66184-1-Ig (same clone as 66184-1-PBS)

  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). This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

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  IHC staining of human liver cancer using 66184-1-Ig (same clone as 66184-1-PBS)

  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). This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

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  IHC staining of human endometrial cancer using 66184-1-Ig (same clone as 66184-1-PBS)

  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). This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

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  IHC staining of human liver cancer using 66184-1-Ig (same clone as 66184-1-PBS)

  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). This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

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  IHC staining of human colon cancer using 66184-1-Ig (same clone as 66184-1-PBS)

  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). This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

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  IF Staining of U2OS using 66184-1-Ig (same clone as 66184-1-PBS)

  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). This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

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  IF Staining of HepG2 using 66184-1-Ig (same clone as 66184-1-PBS)

  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). This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

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  IF Staining of U2OS using 66184-1-Ig (same clone as 66184-1-PBS)

  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). This data was developed using the same antibody clone with 66184-1-PBS in a different storage buffer formulation.

• Featured Product
• KD/KO Validated

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

Cat No. 66184-1-PBS

Clone No.1H5C1

Host / Isotype

Mouse / IgG2b

Reactivity

human

Applications

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

Synonyms

P62,SQSTM1, P62, SQSTM1, 1H5C1, EBI3-associated protein of 60 kDa

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Product Information

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

• Tested Reactivity: human
• 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
• UNIPROT ID: Q13501
• Storage Buffer: PBS only , pH 7.3
• Storage Conditions: Store at -80°C.

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.