Anti-SQSTM1 / p62 antibody - Autophagosome Marker

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (AB155686)

Paraffin-embedded human lung cancer tissue stained for SQSTM1 / P62 protein at cytoplasm with ab155686 at a 1/500 dilution in immunohistochemical analysis.

Antigen Retrieval : Citrate buffer, pH 6.0, 15 min

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (AB155686)

Immunocytochemistry/ Immunofluorescence analysis of HepG2 cells treated with 3μM thapsigargin for 12 hours (right) and mock (left) labeling SQSTM1/p62 with ab155686 at 1 : 500 (green). Cells were fixed in ice-cold MeOH for 10 minutes, permeabilized with cooled acetone for 1 minute. Cells were co-stained with Hoechst (blue).

• Flow Cyt

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Flow Cytometry - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (AB155686)

4% paraformaldehyde-fixed HeLa cells stained for SQSTM1/p62 (blue) using ab155686 (1/100) in Flow Cytometry. The cells were fixed at 4°C for 5 minutes. The brown line is unlabelled sample. Acquisition of >20,000 events were collected using Argon ion laser (488nm) and 525/30 bandpass filter.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (AB155686)

Immunocytochemistry/Immunofluorescent analysis of HeLa cells mock (left) and treated with 50μM Chloroquine for 24 hr (right) labeling SQSTM1/p62 (green) with ab155686 at 1/1000 dilution. Cells were fixed with 4% paraformaldehyde at RT for 15 min. Red : Phalloidin, a F-actin marker.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (AB155686)

ab155686 staining SQSTM1/p62 (autophagosome) in control HeLa cells (left panel) and SQSTM1/p62 in HeLa cells treated with 1uM bafilomycin A1 (ab120497) for 18hrs (right panel). The cells were fixed with methanol (5min), 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 ab155686 at 1/500 dilution and ab7291 (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) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IP

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Immunoprecipitation - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (AB155686)

Immunoprecipitation of SQSTM1/p62 protein from HeLa whole cell extracts using 5 μg of ab155686.
Western blot analysis was performed using ab155686.
Anti-Rabbit IgG was used as a secondary reagent.

All lanes:

Immunoprecipitation - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab155686)

Predicted band size: 47 kDa

false

• WB

Lab

Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (AB155686)

10% SDS-PAGE

All lanes:

Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab155686) at 1/500 dilution

Lane 1:

HepG2 whole cell extracts at 30 µg

Lane 2:

SQSTM1/p62 shRNA transfected HepG2 whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 47 kDa

false

• WB

Supplier Data

Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (AB155686)

Samples were separated by 12% SDS-PAGE.

All lanes:

Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab155686) at 1/1000 dilution

Lane 1:

Untreated Huh-7 whole cell extracts at 30 µg

Lane 2:

Treated Huh-7 whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 48 kDa

false

• WB

Supplier Data

Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (AB155686)

Samples were separated by 10% SDS-PAGE.

All lanes:

Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab155686) at 1/1000 dilution

Lane 1:

Untreated HepG2 whole cell extracts at 30 µg

Lane 2:

Treated HepG2 whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 48 kDa

false

• WB

Supplier Data

Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (AB155686)

10% SDS-PAGE

All lanes:

Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab155686) at 1/1000 dilution

Lane 1:

PC-12 whole cell lysate/extract at 30 µg

Lane 2:

Rat2 whole cell lysate/extract at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 47 kDa

false

• WB

Supplier Data

Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (AB155686)

Samples were separated by 7.5% SDS-PAGE.

All lanes:

Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab155686) at 1/5000 dilution

Lane 1:

Untreated NIH-3T3 whole cell extracts at 30 µg

Lane 2:

Treated NIH-3T3 whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 48 kDa

false

• WB

Supplier Data

Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (AB155686)

Samples were separated by SDS-PAGE.

All lanes:

Western blot - Anti-SQSTM1 / p62 antibody - Autophagosome Marker (ab155686) at 1/1000 dilution

Lane 1:

NIH-3T3 whole cell lysate at 30 µg

Lane 2:

JC whole cell lysate at 30 µg

Lane 3:

BCL-1 whole cell lysate at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 48 kDa

false