Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human

Applications

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Knockout Validated, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit Clone # 2533B

Format

BSA Free

Concentration

1.0 mg/ml

View all p62/SQSTM1 Antibodies »

Product Summary for p62/SQSTM1 Antibody (2533B) - BSA Free

Immunogen

Partial recombinant human p62/SQSTM1 protein produced in E. coli (amino acids 368-440) [UniProt Q13501]

Clonality

Monoclonal

Host

Rabbit

Scientific Data Images for p62/SQSTM1 Antibody (2533B) - BSA Free

Knockout Validated: p62/SQSTM1 Antibody (2533B) - BSA Free [NBP2-92995]

Knockout Validated: p62/SQSTM1 Antibody (2533B) - BSA Free [NBP2-92995]

Knockout Validated: p62/SQSTM1 Antibody (2533B) [NBP2-92995] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and p62/SQSTM1 knockout HeLa cell line (KO). PVDF membrane was probed with 1 ug/mL of Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # NBP2-92995) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for p62/SQSTM1 at approximately 60 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # 2275-PC-100) is shown as a loading control. This experiment was conducted under reducing conditions.
Western Blot: p62/SQSTM1 Antibody (2533B)BSA Free [NBP2-92995]

Western Blot: p62/SQSTM1 Antibody (2533B)BSA Free [NBP2-92995]

Western Blot: p62/SQSTM1 Antibody (2533B) [NBP2-92995] - Western blot shows lysates of A172 human glioblastoma cell line and RT-4 human bladder carcinoma cell line. PVDF membrane was probed with 1 ug/mL of Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # NBP2-92995) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for p62/SQSTM1 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions.
Immunocytochemistry/ Immunofluorescence: p62/SQSTM1 Antibody (2533B) - BSA Free [NBP2-92995]

Immunocytochemistry/ Immunofluorescence: p62/SQSTM1 Antibody (2533B) - BSA Free [NBP2-92995]

Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (2533B) [NBP2-92995] - p62/SQSTM1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with chloroquine (left) or untreated (right) using Rabbit Anti-Human p62/SQSTM1 Monoclonal Antibody (Catalog # NBP2-92995) at 8 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm.
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Applications for p62/SQSTM1 Antibody (2533B) - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

8-25 ug/ml

Immunohistochemistry

8-25 ug/ml

Immunohistochemistry-Paraffin

8-25 ug/ml

Simple Western

20 ug/ml

Western Blot

1 ug/ml
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

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

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. 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 (2533B) - BSA Free

Product Datasheets

Certificate of Analysis

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Product Specific Notices for p62/SQSTM1 Antibody (2533B) - 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.

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