Anti-SQSTM1 / p62 antibody [2C11] - is a mouse monoclonal antibody that is provided in a PBS only formulation free from BSA and azide. Formulated to be conjugation-ready. It is used to detect SQSTM1 / p62 in Flow cytometry, ICC/IF, IHC-P, IP, Western blot. Suitable for Human samples.
- PBS only, conjugation-ready, removing anything extra from your antibodies, greater flexibility in assay design
- Antibody clone 2C11 is the most widely used clone for SQSTM1 / p62 on the market
- Specificity confirmed with SQSTM1 knockout cell line validation
- One antibody for all your SQSTM1 / p62 staining
pH: 7.4
Constituents: PBS
IP | Flow Cyt | WB | IHC-P | ICC/IF | |
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Human | Tested | Tested | Tested | Tested | Tested |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. We recommend Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) secondary antibody |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes Fix cells for 15 minutes with 2 mL of 4% paraformaldehyde solution (pH 7.4 with NaOH in PBS). Permeabilize cells by incubating for 15 minutes on ice with 2 mL of 0.1% Triton X-100 in PBS. |
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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.
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
Anti-SQSTM1 / p62 antibody [2C11] - is a mouse monoclonal antibody that is provided in a PBS only formulation free from BSA and azide. Formulated to be conjugation-ready. It is used to detect SQSTM1 / p62 in Flow cytometry, ICC/IF, IHC-P, IP, Western blot. Suitable for Human samples.
- PBS only, conjugation-ready, removing anything extra from your antibodies, greater flexibility in assay design
- Antibody clone 2C11 is the most widely used clone for SQSTM1 / p62 on the market
- Specificity confirmed with SQSTM1 knockout cell line validation
- One antibody for all your SQSTM1 / p62 staining
pH: 7.4
Constituents: PBS
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Terms & Conditions.
ab56416 was shown to react with SQSTM1/p62 in wild-type HEK293T cells in western blot. Loss of signal was observed when knockout cell line Human SQSTM1 (p62) knockout HEK-293T cell line ab255343 (knockout cell lysate Human SQSTM1 (p62) knockout HEK-293T cell lysate ab263770) was used. Wild-type HEK293T and SQSTM1 knockout HEK293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab56416 and Anti-GAPDH antibody[EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-SQSTM1 / p62 antibody [2C11] - BSA and Azide free (ab56416) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: SQSTM1 knockout HEK293T cell lysate at 20 µg
Lane 2: Western blot - Human SQSTM1 (p62) knockout HEK-293T cell line (Human SQSTM1 (p62) knockout HEK-293T cell line ab255343)
Performed under reducing conditions.
Predicted band size: 47 kDa
Observed band size: 62 kDa
Immunoprecipitation of SQSTM1 in U-2 OS cells. Lysates were prepared and immunoprecipitation was performed using 1.0 μg of ab56416 pre-coupled to prot.G-Sepharose beads. Samples were washed and processed for western blot with Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker ab207305 at 1/10,000. 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 [2C11] - BSA and Azide free (ab56416)
Predicted band size: 47 kDa
Overlay histogram showing HeLa cells stained with ab56416 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56416, 0.5μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (Mouse IgG2a, Kappa Monoclonal [B12/8] - Isotype Control ab91361, 1μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
This image was generated using the ascites version of the product.
ab56416 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 1/10 000. The cells were then incubated with ab56416 at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-mouse 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.ing knock out cell lines.
ab56416 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 ab56416 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with goat anti-mouse HRP secondary antibodies at 0.3ug/mL before imaging. This data was kindly provided by the YCharOS Inc., an open science company with the mission of characterizing every commercially available antibody reagent. Abcam are working with YCharOS to support their mission of antibody characterisation using knockout cell lines.
All lanes: Western blot - Anti-SQSTM1 / p62 antibody [2C11] - BSA and Azide free (ab56416) 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
Monoclonal antibody to SQSTM1 (ab56416) on HeLa cell, antibody concentration 10 ug/ml.
This image was generated using the ascites version of the product.
ab56416 (1μg/ml) staining SQSTM1 in human lymph node using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear and cytoplasmic staining.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
This image was generated using the ascites version of the product.
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