Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635)

Anti-SQSTM1 / p62 antibody [EPR23101-103] (ab240635) is a rabbit monoclonal antibody that is used to detect SQSTM1 / p62 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, ICC/IF. Suitable for Human, Mouse, Rat samples.

- Specificity confirmed with SQSTM1 / p62 knockout cell line validation

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Images

Immunocytochemistry - Anti-SQSTM1 / p62 antibody [EPR23101-103] (AB240635), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	IHC-P	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Not recommended	Tested	Tested
Mouse	Tested	Tested	Not recommended	Tested	Tested
Rat	Expected	Tested	Not recommended	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes -
Species Human	Dilution info 1/30	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1 µg/mL	Notes -
Species Human	Dilution info 1 µg/mL	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -
Species Human	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

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Target data

See full target information SQSTM1

Function

The protein expressed by the SQSTM1 gene is an autophagy receptor essential for selective macroautophagy (aggrephagy). It acts as a bridge between polyubiquitinated cargo and autophagosomes, interacting with both the cargo and an autophagy modifier from the MAP1 LC3 family. Alongside WDFY3, it plays a role in forming and autophagically degrading cytoplasmic ubiquitin-containing inclusions and recruiting ubiquitinated proteins to nuclear PML bodies. SQSTM1 may regulate NFKB1 activation by TNF-alpha, nerve growth factor (NGF), and interleukin-1, and may influence titin/TTN signaling in muscle cells. It may also affect signaling cascades via ubiquitination and be involved in cell differentiation, apoptosis, immune response, and regulation of potassium channels. The protein is involved in endosome organization by retaining vesicles in the perinuclear cloud; ubiquitination by RNF26 allows it to attract vesicle-associated adapters, forming a molecular bridge to organize endosomal pathways. Additionally, it promotes the relocalization of Lys-63-linked ubiquitinated STING1 to autophagosomes and acts as an activator of the NFE2L2/NRF2 pathway by interacting with KEAP1, which inactivates the BCR(KEAP1) complex, resulting in nuclear accumulation of NFE2L2/NRF2 and expression of cytoprotective genes. This supplementary information is collated from multiple sources and compiled automatically.

Alternative names

ORCA, OSIL, SQSTM1, Sequestosome-1, EBI3-associated protein of 60 kDa, Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa, Ubiquitin-binding protein p62, EBIAP, p60, p62

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR23101-103
Purification technique: Affinity purification Protein A
Specificity: WB: This antibody is not suitable for using in rat tissues. We suggest loading 40µg lysate per lane in gel to obtain good signal.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-SQSTM1 / p62 antibody [EPR23101-103] (ab240635) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunocytochemistry/immunofluorescence (ICC/IF), in Human, Mouse, Rat samples.

What is the molecular weight of SQSTM1 / p62?

Anti-SQSTM1 / p62 [EPR23101-103] (ab240635) specifically detects a band for SQSTM1 / p62 (UniProt: Q64337) at a molecular weight of 47kDa.

Recommended positive controls

WB: U-2 OS, HeLa, MEF, NIH/3T3, PC-12, C2C12 and J774A.1 lysates. ICC/IF: HeLa, U-2 OS and MEF cells. Flow Cyt (intra): HeLa and MEF cells. IP: HeLa, U-2 OS and MEF cells.

Trusted by the scientific community

Anti-SQSTM1 / p62 [EPR23101-103] (ab240635) was first used in a scientific publication in 2019 and has been cited over 10 times in peer-reviewed journals.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies. Specificity confirmed

The specificity of Anti-SQSTM1 / p62 antibody [EPR23101-103] (ab240635) has been confirmed by Immunocytochemistry testing in SQSTM1 Knockout HAP1 cells.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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12 product images

Immunocytochemistry - Anti-SQSTM1 / p62 antibody [EPR23101-103] (ab240635)
ab240635 staining SQSTM1 in wild-type Hap1 cells and SQSTM1 knockout Hap1 cells treated with chloroquine (Chloroquine diphosphate, apoptosis and autophagy inhibitor ab142116, 50μM for 24 hrs). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab240635 at 1μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635)
Immunoprecipitation of SQSTM1 in U-2 OS cells. Lysates were prepared and immunoprecipitation was performed using 1.0 μg of ab240635 pre-coupled to prot.A-Sepharose beads. Samples were washed and processed for western blot with Anti-SQSTM1 / p62 antibody [2C11] - BSA and Azide free ab56416 at 1/5000. SM=10% starting material; UB=10% unbound fraction; IP=immunoprecipitate. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635)
Predicted band size: 47 kDa
Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR23101-103] (ab240635)
ab240635 staining SQSTM1/p62 (autophagosome) in control HeLa cells (left panel) and SQSTM1/p62 in HeLa cells treated with 100nM bafilomycin A1 (Bafilomycin A1, (V)-ATPase inhibitor ab120497) for 18hrs (right panel). The cells were fixed with methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab240635 at 2ug/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Western blot - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635)
SQSTM1 / p62 Western blot staining using rabbit Anti-SQSTM1 / p62 antibody
ab240635 was shown to react with SQSTM1 in wild-type U-2 OS cells in Western blot with loss of signal observed in a SQSTM1 knockout cell line. Wild-type U-2 OS and SQSTM1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab240635 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2ug/mL before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635) at 1/1000 dilution
Lane 1: Wild-type U-2 OS cell lysate at 20 µg
Lane 2: SQSTM1 knockout U-2 OS cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 47 kDa
Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635)
SQSTM1 / p62 was immunoprecipitated from 0.35 mg MEF (Mouse embryonic fibroblast (immortalized)) whole cell lysate with ab240635 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab240635 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: MEF whole cell lysate 10ug
Lane 2: ab240635 IP in MEF whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab240635 in MEF whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 min.
All lanes: Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635)
Predicted band size: 47 kDa
Observed band size: 62 kDa
Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR23101-103] (ab240635)
ab240635 was shown to react with SQSTM1 in wild-type U-2 OS cells in Immunocytochemistry with loss of signal observed in a SQSTM1 knockout cell line. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1/5000. The cells were then incubated with ab240635 at 1/500 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor^® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR23101-103] (ab240635)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MEF (Mouse embryo fibroblast) cells labelling SQSTM1 / p62 with ab240635 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in MEF cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.
Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635)
SQSTM1 / p62 was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate with ab240635 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab240635 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HeLa whole cell lysate 10ug
Lane 2: ab240635 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab240635 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
All lanes: Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635)
Predicted band size: 47 kDa
Observed band size: 62 kDa
Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635)
SQSTM1 / p62 Flow Cytometry (Intracellular) staining using rabbit Anti-SQSTM1 / p62 antibody
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized MEF (Mouse embryonic fibroblast (immortalized)) cells labelling SQSTM1 / p62 with ab240635 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635)
SQSTM1 / p62 Western blot staining using rabbit Anti-SQSTM1 / p62 antibody
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times.
Lanes 1-4:3 minutes; Lanes 5-6:92 seconds.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 24086455).
All lanes: Western blot - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635) at 1/1000 dilution
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 2: MEF (mouse embryonic fibroblast (immortalized)), whole cell lysate at 20 µg
Lane 3: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg
Lane 4: PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 20 µg
Lane 5: C2C12 (mouse myoblasts myoblast), whole cell lysate at 20 µg
Lane 6: J774A.1 (mouse reticulum cell sarcoma monocyte macrophage), whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 47 kDa
Observed band size: 62 kDa
Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (ab240635)
SQSTM1 / p62 Flow Cytometry (Intracellular) staining using rabbit Anti-SQSTM1 / p62 antibody
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling SQSTM1 / p62 with ab240635 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR23101-103] (ab240635)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling SQSTM1 / p62 with ab240635 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.