APPLICATION
Application Note
*Optimal dilutions/concentrations should be determined by the researcher.
Application |
Dilution |
1:500-1:3000 |
1:100-1:1000 |
1:100-1:1000 |
1:50-1:200 |
Not tested in other applications.
Calculated MW
Positive Control
HepG2 , A549 , H1299 , HCT116 , Huh-7 (untreated) , Huh-7 (3 μM Thapsigargin treatment for 12 hr) , Huh-7 (3 μM Thapsigargin treatment for 24 hr) , HepG2 (3 μM Thapsigargin treatment for 12 hr) , HepG2 (3 μM Thapsigargin treatment for 24 hr) , *2C4 , *MEF , *MNT-1
PROPERTIES
Form
Liquid
Buffer
PBS
Preservative
No Preservative
Storage
Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles.
Concentration
1 mg/ml (Please refer to the vial label for the specific concentration.)
Antigen Species
Human
Immunogen
Recombinant protein encompassing a sequence within the center region of human SQSTM1 / P62. The exact sequence is proprietary.
Purification
Affinity purified by Protein G.
Conjugation
Unconjugated
RRID
AB_2885144
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET
Synonyms
sequestosome 1 , A170 , DMRV , FTDALS3 , NADGP , OSIL , PDB3 , ZIP3 , p60 , p62 , p62B
Cellular Localization
Cytoplasm , Late endosome , Nucleus
Background
This gene encodes a multifunctional protein that binds ubiquitin and regulates activation of the nuclear factor kappa-B (NF-kB) signaling pathway. The protein functions as a scaffolding/adaptor protein in concert with TNF receptor-associated factor 6 to mediate activation of NF-kB in response to upstream signals. Alternatively spliced transcript variants encoding either the same or different isoforms have been identified for this gene. Mutations in this gene result in sporadic and familial Paget disease of bone. [provided by RefSeq]
Database
Research Area
DATA IMAGES
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GTX629890 WB Image
SQSTM1 antibody detects SQSTM1 protein by western blot analysis. Un-treated (-) and treated (+, Thapsigargin treatment for 12 hr and 24hr) Huh-7 whole cell extracts (30 μg) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody (GTX629890) diluted by 1:1000. The ACTB was used as internal control (GTX110564, 1:50000) shown at the bottom panel. The HRP-conjugated anti-mouse IgG antibody (GTX213111-01) was used to detect the primary antibody.
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GTX629890 WB Image
Various whole cell extracts (30 μg) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 / P62 antibody [GT1478] (GTX629890) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody (GTX213111-01) was used to detect the primary antibody.
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GTX629890 ICC/IF Image
SQSTM1 antibody [GT1478] detects SQSTM1 protein at autophagosome by immunofluorescent analysis. Samples: HeLa cells mock (left) and treated with 50μM Chloroquine for 24 hr (right) were fixed in 4% paraformaldehyde at RT for 15 min. Green: SQSTM1 protein stained by SQSTM1 antibody [GT1478] (GTX629890) diluted at 1:1000. Red: phalloidin, a cytoskeleton marker, stained by phalloidin (invitrogen, A12380) diluted at 1:200. Blue: Hoechst 33342 staining.
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GTX629890 IHC-Fr Image
SQSTM1 antibody [GT1478] detects SQSTM1 protein expression by immunohistochemical analysis. Sample: Frozen sectioned adult mouse retina. Green: SQSTM1 protein stained by SQSTM1 antibody [GT1478] (GTX629890) diluted at 1:250. Red: beta Tubulin 3/ TUJ1, stained by beta Tubulin 3/ TUJ1 antibody [GT11710] (GTX631836) diluted at 1:250. Blue: Fluoroshield with DAPI (GTX30920).
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GTX629890 WB Image
Non-transfected (–) and transfected (+) HepG2 whole cell extracts (50 μg) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody [GT1478] (GTX629890) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody (GTX213111-01) was used to detect the primary antibody.
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GTX629890 FACS Image
SQSTM1 antibody [GT1478] (GTX629890) detects SQSTM1 protein by flow cytometry analysis. Sample: HeLa cell fixed in 4% paraformaldehyde at 4ºC for 5 min. Brown: Unlabelled sample was also used as a control. Blue: SQSTM1 antibody [GT1478] (GTX629890) dilution: 1:100. Acquisition of >20,000 events were collected using Argon ion laser (488nm) and 525/30 bandpass filter.
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GTX629890 ICC/IF Image
SQSTM1 antibody [GT1478] detects SQSTM1 protein at autophagosome by immunofluorescent analysis. Samples: HepG2 cells treated with 3μM thapsigargin 16 hrs (rigtht) and mock (left) were fixed in ice-cold MeOH for 10 min, permeabilize with cooled acetone for 1 min . Green: SQSTM1 protein stained by SQSTM1 antibody [GT1478] (GTX629890) diluted at 1:500. Blue: Hoechst 33342 staining.
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GTX629890 WB Image
SQSTM1 antibody detects SQSTM1 protein by western blot analysis. Un-treated (-) and treated (+, Thapsigargin treatment for 12 hr and 24hr) HepG2 whole cell extracts (30 μg) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody (GTX629890) diluted by 1:1000. The ACTB was used as internal control (GTX110564, 1:50000) shown at the bottom panel. The HRP-conjugated anti-mouse IgG antibody (GTX213111-01) was used to detect the primary antibody.
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REFERENCE
REVIEW
SQSTM1 / P62 antibody [GT1478]
Cat No. GTX629890
Rating
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( Average 4.6 based on 7 users reviews)
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Application
Western Blot(WB)
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( Average 4.6 based on 7 users reviews)
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Date :
Anonymous submitted on 20-May-2021
Application Tested :
WB
Sample Species :
Hu
Sample :
HCE-T cells
Amount used :
2 μg
Blocking :
Blocking One-P, 25°C, 1Hr
Primary Antibody :
1:5000, 4°C, 18Hr
Date :
Anonymous submitted on 06-May-2021
Application Tested :
WB
Sample Species :
Hu
Sample :
A549 cells
Amount used :
10 μg
Blocking :
5% skim milk in TBST, 27°C, 1Hr
Primary Antibody :
1:5000, 4°C, 12Hr
Lane Description :
Two groups (mock and Dengue virus infection) of whole cell extracts (10 μg) were separated by 12% SDS-PAGE, and the membrane was blotted with SQSTM1 / P62 antibody (GTX629890) diluted at 1:5000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
Date :
Anonymous submitted on 24-May-2018
Application Tested :
WB
Sample Species :
Ms
Sample :
Raw264.7 cells
Amount used :
50 μg
Blocking :
5% milk, 25°C, 1Hr
Primary Antibody :
1:1000, 37°C, 1Hr
Date :
Anonymous submitted on 12-May-2016
Application Tested :
WB
Sample Species :
Hu
Sample :
HepG2 cells
Amount used :
30μg
Blocking :
3% BSA in PBST, 25°C, 1Hr
Primary Antibody :
1:1000, 25°C, 16Hr
Date :
Anonymous submitted on 24-Apr-2015
Application Tested :
WB
Sample Species :
Hu
Sample :
MNT-1 cells
Amount used :
30μg
Blocking :
5% skim milk in TBST, 25°C, 1Hr
Primary Antibody :
1:1000, 4°C, 16Hr
Date :
Anonymous submitted on 18-Mar-2015
Application Tested :
WB
Sample Species :
Ms
Sample :
MEF cells
Amount used :
50μg
Blocking :
5% non-fat milk, 25°C, 1Hr
Primary Antibody :
1:1000, 4°C, 14Hr
Date :
Anonymous submitted on 17-Mar-2015
Application Tested :
WB
Sample Species :
Hu
Sample :
2C4 cells
Amount used :
50μg
Blocking :
5% non-fat milk, 25°C, 1Hr
Primary Antibody :
1:1000, 4°C, 14Hr