p62/SQSTM1 Antibody (1478) - Azide and BSA Free

Novus Biologicals | Catalog # NBP2-43663

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Human, Mouse

Applications

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 1478

Format

Azide and BSA Free
View all p62/SQSTM1 Primary Antibodies »

Product Specifications

Immunogen

Recombinant protein encompassing a sequence within the center region of human p62/SQSTM1. The exact sequence is proprietary.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Theoretical MW

48 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for p62/SQSTM1 Antibody (1478) - Azide and BSA Free

Western Blot: p62/SQSTM1 Antibody (1478) [NBP2-43663]

Western Blot: p62/SQSTM1 Antibody (1478) [NBP2-43663]

Western Blot: p62/SQSTM1 Antibody (1478) [NBP2-43663] - Analysis of un-treated (-) and treated (+), Thapsigargin treatment for 12hr and 24hr. HepG2 whole cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody (1478) dilution 1:1000. Beta Actin was used as internal control (bottom panel).
Western Blot: p62/SQSTM1 Antibody (1478) [NBP2-43663]

Western Blot: p62/SQSTM1 Antibody (1478) [NBP2-43663]

Western Blot: p62/SQSTM1 Antibody (1478) [NBP2-43663] - Analysis of un-treated (-) and treated (+), Thapsigargin treatment for 12hr and 24hr. Huh-7 whole cell extracts (30 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody (1478) dilution 1:1000. Beta Actin was used as internal control (bottom panel).
Immunocytochemistry/ Immunofluorescence: p62/SQSTM1 Antibody (1478) [NBP2-43663]

Immunocytochemistry/ Immunofluorescence: p62/SQSTM1 Antibody (1478) [NBP2-43663]

Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (1478) [NBP2-43663] - Analysis of Samples: HeLa cells mock (left) and treated with 50uM Chloroquine for 24 hr (right) were fixed in 4% paraformaldehyde at RT for 15 min. Green: SQSTM1 protein stained by SQSTM1 antibody [1478]. Red: phalloidin, a cytoskeleton marker, stained by phalloidin diluted at 1:200. Blue: Hoechst 33342 staining.
Flow Cytometry: p62/SQSTM1 Antibody (1478) [NBP2-43663]

Flow Cytometry: p62/SQSTM1 Antibody (1478) [NBP2-43663]

Flow Cytometry: p62/SQSTM1 Antibody (1478) [NBP2-43663] - Analysis of HeLa cell. Brown: Unlabelled sample was also used as a control. Blue: SQSTM1 antibody [1478] dilution: 1:100.
p62/SQSTM1 Antibody (1478) - Azide and BSA Free

Western Blot: p62/SQSTM1 Antibody (1478) [NBP2-43663] -

Non-transfected (-) and transfected (+) HepG2 whole cell extracts (50 ug) were separated by 10% SDS-PAGE, and the membrane was blotted with SQSTM1 antibody [GT1478] (NBP2-43663) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
p62/SQSTM1 Antibody (1478) - Azide and BSA Free

Immunohistochemistry-Frozen: p62/SQSTM1 Antibody (1478) [NBP2-43663] -

SQSTM1 antibody [GT1478] detects SQSTM1 protein expression by immunohistochemical analysis.
Sample: Frozen sectioned adult mouse retina.
Green: SQSTM1 protein stained by SQSTM1 antibody [GT1478] (NBP2-43663) diluted at 1:250.
Red: beta Tubulin 3/ TUJ1, stained by beta Tubulin 3/ TUJ1 antibody [GT11710] diluted at 1:250.
Blue: Fluoroshield with DAPI.

Applications for p62/SQSTM1 Antibody (1478) - Azide and BSA Free

Application
Recommended Usage

Flow Cytometry

1:50-1:200

Immunocytochemistry/ Immunofluorescence

1:100-1:1000

Immunohistochemistry-Frozen

1:100-1:1000

Western Blot

1:500-1:3000

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS, 20% Glycerol

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

Concentrations vary lot to lot. See vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: p62/SQSTM1

p62/SQSTM1 (ubiquitin-binding protein p62/Sequestrosome-1) is an intracellular protein (theoretical molecular weight 47.7 kDa) which localizes to the cytoplasm, nucleus, autophagosomes and lysosomes in all tissues (1). Structurally, p62/SQSTM1 consists of multiple domains (e.g., Phox1 and Bem1p-PB1, zinc finger-ZZ, TRAF6 binding domain-TB, Keap1-interacting region-KIR, LC3 interacting region-LIR and ubiquitin-associated domain-UBA) for interaction with various protein targets. Additional domains include nuclear export (NES) and nuclear localization signals (NLS1 and NLS2). p62/SQSTM1 is a multifunctional scaffold protein which forms oligomers with itself or with other proteins through interactions with PB1 domains.

Abnormal function of p62/SQSTM1 is associated with a range of disease states such as neurodegeneration, cancer, and metabolic disorders (2). Mutations in the p62/SQSTM1 sequence have been linked to Paget's disease of the bone, amyotrophic lateral sclerosis, and frontotemporal lobar degeneration. In Parkinson's disease, p62/SQSTM1 has been linked to microglia activation and subsequent neuroinflammation (3). Functionally, p62/SQSTM1 is involved in a broad range of cellular processes such as amino acid sensing by interaction with mTORC1, oxidative stress response through interaction with Keap1, and targeting cargo for autophagy by interacting with ubiquitin labeled proteins (1).

To induce selective autophagy, p62/SQSTM1 forms long oligomers or helical filaments which interact with LC3 and ubiquitin labeled proteins and lead to the initiation of the autophagosome formation (2). p62/SQSTM1 is not only a selective autophagy receptor but also an autophagy substrate, as its engulfed by the autophagosome and degraded by the autophagolysosome. Monitoring LC3 levels is the standard for assessing autophagic flux, however monitoring p62/SQSTM1 levels by Western blot in the presence and absence of autophagy inhibitors (e.g., Chloroquine) is also a common practice (4). Besides its activity as a selective autophagy receptor, p62/SQSTM1 also plays a role as an adaptor in signaling cascades leading to NFkB activation downstream of TNF-R, IL-1 beta R, TrkA and p75NTR. Briefly, for NFkB signaling downstream of the TNF-R activation, p62/SQSTM1 engages RIP1 kinase and PKC iota/lambda through the ZZ and PB1 domains, respectively (5).

References

1.Katsuragi, Y., Ichimura, Y., & Komatsu, M. (2015). P62/SQSTM1 functions as a signaling hub and an autophagy adaptor. FEBS Journal. https://doi.org/10.1111/febs.13540

2. Sanchez-Martin, P., & Komatsu, M. (2018). p62/SQSTM1 - Steering the cell through health and disease. Journal of Cell Science. https://doi.org/10.1242/jcs.222836

3. Yao, L., Zhu, Z., Wu, J., Zhang, Y., Zhang, H., Sun, X.,... Lu, G. (2019). MicroRNA-124 regulates the expression of p62/p38 and promotes autophagy in the inflammatory pathogenesis of Parkinson's disease. The FASEB Journal. https://doi.org/10.1096/fj.201900363r

4 Klionsky, D. J., Abdelmohsen, K., Abe, A., Abedin, M. J., Abeliovich, H., Arozena, A. A.,... Zughaier, S. M. (2016). Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. https://doi.org/10.1080/15548627.2015.1100356

5. Bitto, A., Lerner, C. A., Nacarelli, T., Crowe, E., Torres, C., & Sell, C. (2014). p62/SQSTM1 at the interface of aging, autophagy, and disease. Age. https://doi.org/10.1007/s11357-014-9626-3

Long Name

Sequestosome 1

Alternate Names

A170, EBIAP, ORCA, OSIL, PDB3, Sequestosome 1, SQSTM1, ZIP3

Gene Symbol

SQSTM1

Additional p62/SQSTM1 Products

Product Documents for p62/SQSTM1 Antibody (1478) - Azide and BSA Free

Certificate of Analysis

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Product Specific Notices for p62/SQSTM1 Antibody (1478) - Azide and BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Autophagy
NFkB Pathway

Citations for p62/SQSTM1 Antibody (1478) - Azide and BSA Free

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Protocols

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