GeneTex
United States (US)

SQSTM1 / P62 antibody [N3C1], Internal

KO/KD Orthogonal
GTX100685_41052_20141219_IFA_19011506_562.jpg
GTX100685_41052_20150212_FACS_19011506_445.jpg
GTX100685_41052_20150209_IP_19011506_670.jpg
GTX100685_41052_20141226_IHC_19011506_834.jpg
GTX100685_39988_WB_19011506_477.jpg
GTX100685_42824_20170504_WB_Thapsigargin_19011506_430.jpg
GTX100685_39988_WB_M_19011506_981.jpg
GTX100685_41052_20160804_WB_shRNA_watermark_19011506_930.jpg
GTX100685_41052_20160331_WB_thapsigargin_19011506_926.jpg
GTX100685_39988_WB_R_19011506_878.jpg
GTX100685_39988_IFA_19011506_709.jpg

Cat No. GTX100685

Host Rabbit
Clonality Polyclonal
Isotype IgG
Application WB, ICC/IF, IHC-P, FACS, IP
Reactivity Human, Mouse, Rat, Zebrafish, Bovine, Honeybee
APPLICATION

Application Note

*Optimal dilutions/concentrations should be determined by the researcher.
Application Dilution
WB 1:500-1:3000
ICC/IF 1:100-1:1000
IHC-P 1:100-1:1000
FACS 1:50-1:200
IP 1:100-1:500
Not tested in other applications.

Calculated MW

48 kDa. ( Note )

Positive Control

A549 , HepG2 , HepG2 (3μM Thapsigargin treatment for 24 hr) , Huh7 , Huh7 ( 3μM Thapsigargin, 12hr) , NIH-3T3 , JC , BCL-1 , PC-12 , Rat2 , *HeLa

Predict Reactivity

Rhesus Monkey(>80% identity)
PROPERTIES

Form

Liquid

Buffer

1XPBS, 1% BSA, 20% Glycerol (pH7). 0.025% ProClin 300 was added as a 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

0.29mg/ml(Please refer to the vial label for the specific concentration.)

Antigen Species

Human

Immunogen

Carrier-protein conjugated synthetic peptide encompassing a sequence within the center region of human SQSTM1. The exact sequence is proprietary.

Purification

Purified by antigen-affinity chromatography.

Conjugation

Unconjugated

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

A170 antibody, OSIL antibody, PDB3 antibody, ZIP3 antibody, p60 antibody, p62 antibody, p62B antibody, SQSTM1 antibody, EBI3-associated protein p60 antibody, ubiquitin-binding protein p62 antibody, EBI3-associated protein of 60 kDa antibody, oxidative stress induced like antibody, phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa antibody, EBIAP antibody, phosphotyrosine independent ligand for the Lck SH2 domain p62 antibody, sequestosome-1 antibody, sequestosome 1 antibody

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
GTX100685_41052_20141219_IFA_19011506_562.jpg

GTX100685 ICC/IF Image

Orthogonal

SQSTM1 antibody [N3C1], Internal 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 [N3C1], Internal (GTX100685) diluted at 1:1000.
Red: Phalloidin, a F-actin marker.

GTX100685_41052_20150212_FACS_19011506_445.jpg

GTX100685 FACS Image

SQSTM1 antibody [N3C1], Internal (GTX100685) 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 [N3C1], Internal] (GTX100685) dilution: 1:100.
Acquisition of >20,000 events were collected using Argon ion laser (488nm) and 525/30 bandpass filter.

GTX100685_41052_20150209_IP_19011506_670.jpg

GTX100685 IP Image

Immunoprecipitation of SQSTM1 protein from HeLa whole cell extracts using 5 μg of SQSTM1 antibody [N3C1], Internal (GTX100685).
Western blot analysis was performed using SQSTM1 antibody [N3C1], Internal (GTX100685).
EasyBlot anti-Rabbit IgG (GTX221666-01) was used as a secondary reagent.

GTX100685_41052_20141226_IHC_19011506_834.jpg

GTX100685 IHC-P Image

SQSTM1 antibody [N3C1], Internal detects SQSTM1 protein at cytoplasm on human ovarian carcinoma by immunohistochemical analysis.
Sample: Paraffin-embedded human ovarian carcinoma.
SQSTM1 antibody [N3C1], Internal (GTX100685) diluted at 1:500.

Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min

GTX100685_39988_WB_19011506_477.jpg

GTX100685 WB Image

Sample (30 µg of whole cell lysate)
A: A549
10% SDS PAGE
GTX100685 diluted at 1:1000
The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX100685_42824_20170504_WB_Thapsigargin_19011506_430.jpg

GTX100685 WB Image

Untreated (–) and treated (+) HepG2 whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody [N3C1], Internal (GTX100685) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX100685_39988_WB_M_19011506_981.jpg

GTX100685 WB Image

SQSTM1 antibody [N3C1], Internal detects SQSTM1 protein by western blot analysis.
A. 30 µg NIH-3T3 whole cell lysate/extract
B. 30 µg JC whole cell lysate/extract
C. 30 µg BCL-1 whole cell lysate/extract
12% SDS-PAGE
SQSTM1 antibody [N3C1], Internal (GTX100685) dilution: 1:1000
The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX100685_41052_20160804_WB_shRNA_watermark_19011506_930.jpg

GTX100685 WB Image

KO/KD

Non-transfected (–) and transfected (+) HepG2 whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody [N3C1], Internal (GTX100685) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX100685_41052_20160331_WB_thapsigargin_19011506_926.jpg

GTX100685 WB Image

Orthogonal

Untreated (–) and treated (+) Huh-7 whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody [N3C1], Internal (GTX100685) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX100685_39988_WB_R_19011506_878.jpg

GTX100685 WB Image

SQSTM1 antibody [N3C1], Internal detects SQSTM1 protein by western blot analysis.
A. 30 µg PC-12 whole cell lysate/extract
B. 30 µg Rat2 whole cell lysate/extract
10% SDS-PAGE
SQSTM1 antibody [N3C1], Internal (GTX100685) dilution: 1:1000
The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

GTX100685_39988_IFA_19011506_709.jpg

GTX100685 ICC/IF Image

Orthogonal

SQSTM1 antibody [N3C1], Internal detects SQSTM1 protein at autophagosome by immunofluorescent analysis.
Samples: HepG2 cells treated with 3μM thapsigargin 12 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 [N3C1], Internal (GTX100685) diluted at 1:500.
Blue: Hoechst 33342 staining.
Scale bar = 10 μm.

REFERENCE
REVIEW

SQSTM1 / P62 antibody [N3C1], Internal Cat No. GTX100685

Rating ( Average 4 based on 2 users reviews)
Application
Western Blot(WB) ( Average 4 based on 2 users reviews)
Date : Anonymous submitted on 17-Mar-2015
Score :
Application Tested : WB
Sample Species : Hu
Sample : A549 cells
Amount used : 50μg
Blocking : 5% non-fat milk, 4°C, 14Hr
Primary Antibody : 1:1000, 25°C, 1Hr
Date : Anonymous submitted on 26-Jan-2015
Score :
Application Tested : WB
Sample Species : Hu
Sample : HeLa cells
Amount used : 30μg
Blocking : 5% non-fat milk, 25°C, 1Hr
Primary Antibody : 1:1000, 4°C, 16Hr
Application Reference
SDS
PBS.pdf
Glycerol.pdf
Proclin.pdf