Anti-Abi-1 antibody

• ICC

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Immunocytochemistry - Anti-Abi-1 antibody (AB65828)

Immunofluorescence analysis of paraffin-embedded section of human intestine cancer tissue staining ABI1 with ab65828 at 5 µg/mL concentration. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). Blocking was done with 10% goat serum and the tissue section was then incubated with the primary antibody overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1 : 500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI.

• IHC

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Immunohistochemistry - Anti-Abi-1 antibody (AB65828)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of paraffin-embedded sections of human lung cancer tissue staining ABI1 with ab65828 at 2 µg/ml concentration. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). Blocking was done with 10% goat serum and the tissue section was then incubated with the primary antibody overnight at 4°C. The secondary antibody used was Peroxidase Conjugated Goat Anti-rabbit IgG incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

• Flow Cyt

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Flow Cytometry - Anti-Abi-1 antibody (AB65828)

Flow cytometry analysis staining 4% paraformaldehyde-fixed permeabilized MCF-7 cells with ab65828 (Blue) at 1 µg/1x106 cells concentration. Blocking was done using 10% normal goat serum, and the cells were incubated with the primary antibody for 30 minutes at 20°C. The secondary used was DyLight®488 conjugated goat anti-rabbit IgG at 5-10 µg/1x106 cells, and the isotype control was rabbit IgG (Green) at 1 µg/1x10^6 for 30 minutes at 20°C. Control used was unlabelled sample (Red).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Abi-1 antibody (AB65828)

Immunofluorescence analysis of paraffin-embedded section of human breast cancer tissue staining ABI1 with ab65828 at 5 µg/mL concentration. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). Blocking was done with 10% goat serum and the tissue section was then incubated with the primary antibody overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1 : 500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI.

• ICC

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Immunocytochemistry - Anti-Abi-1 antibody (AB65828)

Immunofluorescence analysis of section of U2OS cells staining ABI1 with ab65828 at 5 µg/mL concentration. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. Blocking was done with 10% goat serum and incubated with the primary antibody overnight at 4°C. Secondary antibody Cy3 Conjugated Goat Anti-Rabbit IgG was used at 1/500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI.

• IHC

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Immunohistochemistry - Anti-Abi-1 antibody (AB65828)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of paraffin-embedded section of human colorectal adenocarcinoma tissue staining ABI1 with ab65828 at 2 µg/ml concentration. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). Blocking was done with 10% goat serum and the tissue section was incubated with the pimary antibody overnight at 4°C. The secondary antibody was Peroxidase Conjugated Goat Anti-rabbit IgG incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

• WB

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Western blot - Anti-Abi-1 antibody (AB65828)

Western blot analysis of ABI1 using ab65828 Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 µg of sample under reducing conditions. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with ab65828 at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1/5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for ABI1 at approximately 60-65 kDa. The expected band size for ABI1 is at 55 kDa.

All lanes:

Western blot - Anti-Abi-1 antibody (ab65828)

Lane 1:

Human 293T whole cell lysates at 30 µg

Lane 2:

Human MCF-7 whole cell lysates at 30 µg

Lane 3:

Human U87 whole cell lysates at 30 µg

Lane 4:

Rat brain tissue lysates at 30 µg

Lane 5:

Rat C6 whole cell lysates at 30 µg

Lane 6:

Mouse brain tissue lysates at 30 µg

Lane 7:

Mouse Neuro-2a whole cell lysates at 30 µg

Lane 8:

Mouse C2C12 whole cell lysates at 30 µg

Secondary

All lanes:

Goat anti-rabbit IgG-HRP secondary antibody at 1/5000 dilution

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