Anti-SQSTM1 / p62 antibody [EPR23101-103] - BSA and Azide free

10 product images

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  Immunocytochemistry - Anti-SQSTM1 / p62 antibody [EPR23101-103] - BSA and Azide free (ab269458)

  SQSTM1 / p62 Immunocytochemistry staining using rabbit Anti-SQSTM1 / p62 antibody

  This data was developed using the same antibody clone in a different buffer formulation (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635). Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 staining SQSTM1 in wild-type Hap1 cells and SQSTM1 knockout Hap1 cells treated with chloroquine (Chloroquine diphosphate, apoptosis and autophagy inhibitor ab142116, 50μM for 24 hrs). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 at 1μg/ml concentration and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
  Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

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  Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR23101-103] - BSA and Azide free (ab269458)

  This data was developed using the same antibody clone in a different buffer formulation (ab269458). Immunoprecipitation of SQSTM1 in U-2 OS cells. Lysates were prepared and immunoprecipitation was performed using 1.0 μg of Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 pre-coupled to prot.A-Sepharose beads. Samples were washed and processed for western blot with Anti-SQSTM1 / p62 antibody [2C11] - BSA and Azide free ab56416 at 1/5000. 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 [EPR23101-103] - Autophagosome Marker (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635)

  Predicted band size: 47 kDa

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  Western blot - Anti-SQSTM1 / p62 antibody [EPR23101-103] - BSA and Azide free (ab269458)

  SQSTM1 / p62 Western blot staining using rabbit Anti-SQSTM1 / p62 antibody

  This data was developed using the same antibody in a different buffer formulation (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635).
  

  Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 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 Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 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.

  All lanes: Western blot - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635) 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

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  Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR23101-103] - BSA and Azide free (ab269458)

  SQSTM1 / p62 was immunoprecipitated from 0.35 mg MEF (Mouse embryonic fibroblast (immortalized)) whole cell lysate with Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: MEF whole cell lysate 10ug

  Lane 2: Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 IP in MEF whole cell lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 in MEF whole cell lysate

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  Exposure time: 3 min.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635).

  All lanes: Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635)

  Predicted band size: 47 kDa

  Observed band size: 62 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR23101-103] - BSA and Azide free (ab269458)

  SQSTM1 / p62 Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-SQSTM1 / p62 antibody

  This data was developed using the same antibody clone in a different buffer formulation (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635). Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 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 1/5000. The cells were then incubated with Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 at 1/500 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit 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.

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  Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR23101-103] - BSA and Azide free (ab269458)

  SQSTM1 / p62 Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-SQSTM1 / p62 antibody

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MEF (Mouse embryo fibroblast) cells labelling SQSTM1 / p62 with Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in MEF cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
  

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635).

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  Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR23101-103] - BSA and Azide free (ab269458)

  SQSTM1 / p62 was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate with Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: HeLa whole cell lysate 10ug

  Lane 2: Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 IP in HeLa whole cell lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 in HeLa whole cell lysate

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  Exposure time: 30 seconds

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635).

  All lanes: Immunoprecipitation - Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635)

  Predicted band size: 47 kDa

  Observed band size: 62 kDa

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  Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 antibody [EPR23101-103] - BSA and Azide free (ab269458)

  SQSTM1 / p62 Flow Cytometry (Intracellular) staining using rabbit Anti-SQSTM1 / p62 antibody

  Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized MEF (Mouse embryonic fibroblast (immortalized)) cells labelling SQSTM1 / p62 with Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635).

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  Flow Cytometry (Intracellular) - Anti-SQSTM1 / p62 antibody [EPR23101-103] - BSA and Azide free (ab269458)

  SQSTM1 / p62 Flow Cytometry (Intracellular) staining using rabbit Anti-SQSTM1 / p62 antibody

  Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling SQSTM1 / p62 with Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody [EPR23101-103] - BSA and Azide free (ab269458)

  SQSTM1 / p62 Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-SQSTM1 / p62 antibody

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling SQSTM1 / p62 with Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
  

  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-SQSTM1 / p62 antibody [EPR23101-103] - Autophagosome Marker ab240635).