Anti-ATG12 antibody

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG12 antibody (AB155589)

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue stained for ATG12 protein at cytoplasm with ab155589 at a 1/500 dilution.

Antigen Retrieval : Citrate buffer, pH 6.0, 15 min.

• IP

Supplier Data

Immunoprecipitation - Anti-ATG12 antibody (AB155589)

Immunoprecipitation of Atg 12-Atg5 protein from 293T whole cell extracts using 5 µg of ATG12 antibody.
Western blot analysis was performed using ATG12 antibody at a 1/1000 dilution.

All lanes:

Immunoprecipitation - Anti-ATG12 antibody (ab155589) at 1/1000 dilution

Lane 1:

293T whole cell extracts (input)

Lane 2:

293T whole cell extracts with control (-)

Lane 3:

293T whole cell extracts (+)

Secondary

All lanes:

Anti-Rabbit IgG Antibody

Observed band size: 55 kDa

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• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG12 antibody (AB155589)

Immunohistochemical analysis of paraffin-embedded rat brain tissue stained for ATG12 protein at cytoplasm with ab155589 at a 1/500 dilution.

Antigen Retrieval : Citrate buffer, pH 6.0, 15 min.

• WB

Supplier Data

Western blot - Anti-ATG12 antibody (AB155589)

Samples were separated by 10% SDS-PAGE.

All lanes:

Western blot - Anti-ATG12 antibody (ab155589) at 1/500 dilution

Lane 1:

Non-transfected 293T whole cell extracts at 30 µg

Lane 2:

shRNA Transfected 293T whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG

Observed band size: 55 kDa

false

• WB

Lab

Western blot - Anti-ATG12 antibody (AB155589)

False colour image of Western blot : Anti-ATG12 antibody staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab155589 was shown to bind specifically to ATG12. A band likely to be the unfunctional complex with ATG5 was observed at 52 kDa in wild-type THP-1 cell lysates with no signal observed at this size in Atg12 knockout cell line ab277831 (knockout cell lysate ab278183) - unconjugated functional form not observed at 15 kD. To generate this image, wild-type and Atg12 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-ATG12 antibody (ab155589) at 1/500 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

ATG12 knockout THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human ATG12 knockout THP-1 cell line (<a href='/en-us/products/cell-lines/human-atg12-knockout-thp-1-cell-line-ab277831'>ab277831</a>)

Lane 3:

HCT 116 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 15 kDa

Observed band size: 52 kDa

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• WB

Supplier Data

Western blot - Anti-ATG12 antibody (AB155589)

Samples were separated by 12% SDS-PAGE.

All lanes:

Western blot - Anti-ATG12 antibody (ab155589) at 1/1000 dilution

Lane 1:

293T whole cell extracts at 30 µg

Lane 2:

A431 whole cell extracts at 30 µg

Lane 3:

HeLa whole cell extracts at 30 µg

Lane 4:

HepG2 whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG

Observed band size: 55 kDa

false

• WB

Unknown

Western blot - Anti-ATG12 antibody (AB155589)

12% SDS PAGE

All lanes:

Western blot - Anti-ATG12 antibody (ab155589) at 1/1000 dilution

All lanes:

Rat brain whole cell lysate at 50 µg

Predicted band size: 15 kDa

false

• WB

Unknown

Western blot - Anti-ATG12 antibody (AB155589)

12% SDS PAGE

All lanes:

Western blot - Anti-ATG12 antibody (ab155589) at 1/1000 dilution

All lanes:

NIH3T3 whole cell lysate at 30 µg

Predicted band size: 15 kDa

false