Recombinant Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free (ab280091)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [3/P62 LCK LIGAND] to SQSTM1 / p62 - BSA and Azide free
- Suitable for: IP, IHC-P, WB, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] - BSA and Azide free
See all SQSTM1 / p62 primary antibodies -
Description
Mouse monoclonal [3/P62 LCK LIGAND] to SQSTM1 / p62 - BSA and Azide free -
Host species
Mouse -
Tested applications
Suitable for: IP, IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type HAP1; HeLa; MCF7; HEK-293 whole cell lysates. IHC-P: Human stomach carcinoma, lung carcinoma tissue. ICC/IF: HeLa cells. IP: HeLa whole cell lysate.
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General notes
ab280091 is the carrier-free version of ab280086.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
3/P62 LCK LIGAND -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Dyes/Markers
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab280091 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 |
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 47 kDa.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 47 kDa. |
ICC/IF
Use at an assay dependent concentration. |
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
- Omim: 601530 Human
- SwissProt: Q13501 Human
- Unigene: 709030 Human
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Alternative names
- A170 antibody
- DMRV antibody
- EBI 3 associated protein of 60 kDa antibody
see all
Images
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All lanes : Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] (ab280086) at 1/1000 dilution
Lane 1 : Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell line), whole cell lysate
Lane 2 : SQSTM1 knockout HAP1 (human chronic myelogenous leukemia near-haploid cell line), whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 4 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution
Predicted band size: 47 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?This data was developed using ab280086, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
Lanes 1 - 4: Merged signal (red and green). Green - ab280086 observed at 62 kDa. Red - loading control ab181602 (Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.
Lanes 1-2: ab280086 Anti-SQSTM1/p62 antibody was shown to react with SQSTM1 in HAP1 cells in Western blot. Loss of signal was observed when SQSTM1 knockout sample was used. Wild-type and SQSTM1 knockout samples were subjected to SDS-PAGE. ab280086 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated at 4°C overnight at 1/1000 dilution and 1/20000 dilution respectively.
Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) preadsorbed (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-SQSTM1 / p62 antibody [3/P62 LCK LIGAND] (ab280086) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell ), whole cell lysate
Lane 2 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Predicted band size: 47 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?This data was developed using ab280086, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 24086455).
Lysates were made freshly and used in WB immediately to minimize protein degradation.
Exposure time: 15 seconds
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This data was developed using ab280086 the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human stomach carcinoma tissue labeling SQSTM1 / p62 with ab280086 at 1/1000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human stomach carcinoma. The section was incubated with ab280086 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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This data was developed using ab280086 the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling SQSTM1 / p62 with ab280086 at 1/1000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human lung carcinoma. The section was incubated with ab280086 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
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This data was developed using ab280086 the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling SQSTM1 / p62 with ab280086 at 1/50 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cell line. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594)(Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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This data was developed using ab280086 the same antibody clone in a different buffer formulation.
SQSTM1 / p62 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell ), whole cell lysate 10 ug with ab280086 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280086 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell ), whole cell lysate 10 ug
Lane 2: ab280086 IP in HeLa whole cell lysate
Lane 3: Mouse monoclonal IgG1(ab18443) instead of ab280086 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 26 seconds
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (0)
ab280091 has not yet been referenced specifically in any publications.