Validation Data Gallery

Filter: AllWBIPIHCIF

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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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.
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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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.
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.
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.
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.
×
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.
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.
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.
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.
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.
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.
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 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.
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 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.
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 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.

Featured Product
KD/KO Validated

P62,SQSTM1 Monoclonal antibody, PBS Only

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

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

Formulation: PBS Only

PBS and Azide

PBS Only

Conjugate: Unconjugated

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information

データシート［PDF］を印刷

MSDS

Tested Applications

Recommended dilution

Application	Dilution
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

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 相同性解析による交差性が予測される生物種
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
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.

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

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

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

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

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

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

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

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

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

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