Recombinant Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
![Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)](https://www.abcam.com/ps/products/211/ab211324/Images/ab211324-469617-anti-sqstm1-p62-phospho-s349-antibody-epr20451-western-blot-wildtype-u2os-sqstm1-knockout-u2os-human.jpg "Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20451] to SQSTM1 / p62 (phospho S349)
- Suitable for: Flow Cyt (Intra), ICC/IF, IP, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451]
See all SQSTM1 / p62 primary antibodies -
Description
Rabbit monoclonal [EPR20451] to SQSTM1 / p62 (phospho S349) -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IP, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: U-2 OS, HeLa, C6 and NIH/3T3 whole cell lysates treated with 2µM MG-132 (ab141003) for 18 hours. ICC/IF: HeLa cells treated with 2µM MG-132 (ab141003) for 18 hours. Flow Cyt (intra): HeLa cells treated with 2µM MG-132 (ab141003) for 18 hours. IP: HeLa treated with 2µM MG-132 (ab141003) for 18h whole cell lysate.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20451 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab211324 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Flow Cyt (Intra) |
1/500.
|
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| ICC/IF | (1) |
1/100.
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| IP |
1/30.
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| WB | (2) |
1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 47 kDa).
|
| Notes |
|---|
|
Flow Cyt (Intra)
1/500. |
|
ICC/IF
1/100. |
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IP
1/30. |
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WB
1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 47 kDa). |
Target
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Function
Adapter protein which binds ubiquitin and may regulate the activation of NFKB1 by TNF-alpha, nerve growth factor (NGF) and interleukin-1. May play a role in titin/TTN downstream signaling in muscle cells. May regulate signaling cascades through ubiquitination. Adapter that mediates the interaction between TRAF6 and CYLD (By similarity). May be involved in cell differentiation, apoptosis, immune response and regulation of K(+) channels. -
Tissue specificity
Ubiquitously expressed. -
Involvement in disease
Defects in SQSTM1 are a cause of Paget disease of bone (PDB) [MIM:602080]. PDB is a metabolic bone disease affecting the axial skeleton and characterized by focal areas of increased and disorganized bone turn-over due to activated osteoclasts. Manifestations of the disease include bone pain, deformity, pathological fractures, deafness, neurological complications and increased risk of osteosarcoma. PDB is a chronic disease affecting 2 to 3% of the population above the age of 40 years. -
Sequence similarities
Contains 1 OPR domain.
Contains 1 UBA domain.
Contains 1 ZZ-type zinc finger. -
Domain
The UBA domain binds specifically 'Lys-63'-linked polyubiquitin chains of polyubiquitinated substrates. Mediates the interaction with TRIM55.
The OPR domain mediates homooligomerization and interactions with PRKCZ, PRKCI, MAP2K5 and NBR1.
The ZZ-type zinc finger mediates the interaction with RIPK1. -
Post-translational
modificationsPhosphorylated. May be phosphorylated by PRKCZ (By similarity). Phosphorylated in vitro by TTN. -
Cellular localization
Cytoplasm. Late endosome. Nucleus. Sarcomere (By similarity). In cardiac muscles localizes to the sarcomeric band (By similarity). Localizes to late endosomes. May also localize to the nucleus. Accumulates in neurofibrillary tangles and in Lewy bodies of neurons from individuals with Alzheimer and Parkinson disease respectively. Enriched in Rosenthal fibers of pilocytic astrocytoma. In liver cells, accumulates in Mallory bodies associated with alcoholic hepatitis, Wilson disease, indian childhood cirrhosis and in hyaline bodies associated with hepatocellular carcinoma. - Information by UniProt
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Database links
- Entrez Gene: 8878 Human
- Entrez Gene: 18412 Mouse
- Entrez Gene: 113894 Rat
- Omim: 601530 Human
- SwissProt: Q13501 Human
- SwissProt: Q64337 Mouse
- SwissProt: O08623 Rat
- Unigene: 709030 Human
see all -
Alternative names
- A170 antibody
- DMRV antibody
- EBI 3 associated protein of 60 kDa antibody
see all
Images
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Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution
Lane 1 : Wild-type U-2 OS cell lysate
Lane 2 : SQSTM1 knockout U-2 OS cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 47 kDaab211324 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 ab211324 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.
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![Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)](https://www.abcam.com/ps/products/211/ab211324/Images/ab211324-273940-anti-sqstm1-p62-phospho-s349-antibody-epr20451-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 2μM MG-132 (ab141003) for 18 hours or untreated, labeling SQSTM1 / p62 (phospho S349) with ab211324 at 1/100 dilution, followed by Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) secondary antibody at 1/200 dilution (green).
Confocal image showing cytoplasmic staining on HeLa cell line. The expression increased after treatment with 2μM MG-132 (ab141003) for 18 hours.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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![Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)](https://www.abcam.com/ps/products/211/ab211324/Images/ab211324-273938-anti-sqstm1-p62-phospho-s349-antibody-epr20451-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 2µM MG-132 (ab141003) for 18 hours (red) or untreated (green), labeling SQSTM1 / p62 (phospho S349) with ab211324 at 1/500 dilution compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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![Immunoprecipitation - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)](https://www.abcam.com/ps/products/211/ab211324/Images/ab211324-273936-anti-sqstm1-p62-phospho-s349-antibody-epr20451-immunoprecipitation.jpg)
Immunoprecipitation - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)SQSTM1 / p62 (phospho S349) was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 2μM MG-132 (ab141003) for 18h with ab211324 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab211324 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Lane 1: HeLa treated with 2μM MG-132 (ab141003) for 18h whole cell lysate, 10 µg (Input).
Lane 2: ab211324 IP in HeLa treated with 2μM MG-132 (ab141003) for 18h whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab211324 in HeLa treated with 2μM MG-132 (ab141003) for 18h whole cell lysate.Blocking and dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)](https://www.abcam.com/ps/products/211/ab211324/Images/ab211324-273950-anti-sqstm1-p62-phospho-s349-antibody-epr20451-western-blot.jpg)
Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours
Lane 3 : HeLa whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours, then treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 47 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking/dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)](https://www.abcam.com/ps/products/211/ab211324/Images/ab211324-273947-anti-sqstm1-p62-phospho-s349-antibody-epr20451-western-blot.jpg)
Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution
Lane 1 : Untreated C6 (Rat glial tumor cell line) whole cell lysate
Lane 2 : C6 whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 47 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
-
![Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)](https://www.abcam.com/ps/products/211/ab211324/Images/ab211324-273944-anti-sqstm1-p62-phospho-s349-antibody-epr20451-western-blot.jpg)
Western blot - Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)All lanes : Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324) at 1/1000 dilution
Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 2 : NIH/3T3 whole cell lysate treated with 2µM MG-132 (ab141003) for 18 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 47 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?
Exposure time: 4 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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![Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)](https://www.abcam.com/ps/products/211/ab211324/Images/ab211324-8-benefits-of-recombinant-antibodies.png)
Anti-SQSTM1 / p62 (phospho S349) antibody [EPR20451] (ab211324)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (16)
ab211324 has been referenced in 16 publications.
- Deng Q et al. A detrimental role of NLRP6 in host iron metabolism during Salmonella infection. Redox Biol 49:102217 (2022). PubMed: 34942528
- Li Y et al. Inhibition of CISD2 promotes ferroptosis through ferritinophagy-mediated ferritin turnover and regulation of p62-Keap1-NRF2 pathway. Cell Mol Biol Lett 27:81 (2022). PubMed: 36180832
- Jin R et al. Melatonin Type 2 Receptor Activation Regulates Blue Light Exposure-Induced Mouse Corneal Epithelial Damage by Modulating Impaired Autophagy and Apoptosis. Int J Mol Sci 23:N/A (2022). PubMed: 36232639
- Xu Q et al. Adenosine 5'-monophosphate-activated protein kinase ameliorates bovine adipocyte oxidative stress by inducing antioxidant responses and autophagy. J Dairy Sci 104:4516-4528 (2021). PubMed: 33551161
- Guo J et al. Bone Marrow-Derived Mesenchymal Stem Cells Ameliorate Sepsis-Induced Acute Kidney Injury by Promoting Mitophagy of Renal Tubular Epithelial Cells via the SIRT1/Parkin Axis. Front Endocrinol (Lausanne) 12:639165 (2021). PubMed: 34248837