Anti-Paxillin antibody - C-terminal

• ICC

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Immunocytochemistry - Anti-Paxillin antibody - C-terminal (AB191007)

Immunocytochemical analysis of HeLa cells, labeling Paxillin with ab191007 at 1 μg/ml.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Paxillin antibody - C-terminal (AB191007)

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue, labeling Paxillin with ab191007 at 1 μg/ml.

• Flow Cyt

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Flow Cytometry - Anti-Paxillin antibody - C-terminal (AB191007)

Overlay histogram showing A549 cells stained with PA1804 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with ab191007 1µg/1x10^6 cells for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG 5-10µg/1x10^6 cells was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1µg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

• ICC

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Immunocytochemistry - Anti-Paxillin antibody - C-terminal (AB191007)

PXN was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 µg/mL ab191007 overnight at 4°C. Biotin conjugated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Cy3 Conjugated Avidin. The section was counterstained with DAPI.

• ICC

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Immunocytochemistry - Anti-Paxillin antibody - C-terminal (AB191007)

PXN was detected in immunocytochemical section of PC-3 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5µg/mL ab191007 overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1/100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Paxillin antibody - C-terminal (AB191007)

Immunohistochemical analysis of frozen rat kidney tissue, labeling Paxillin with ab191007 at 1 μg/ml.

• WB

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Western blot - Anti-Paxillin antibody - C-terminal (AB191007)

Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. 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 ab191007at 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 PXN at approximately 65 kDa. The expected band size for PXN is at 65 kDa.

All lanes:

Western blot - Anti-Paxillin antibody - C-terminal (ab191007) at 0.5 µg/mL

Lane 1:

human Hela whole cell lysates, at 30 µg

Lane 2:

human 293T whole cell lysates, at 30 µg

Lane 3:

human HepG2 whole cell lysates, at 30 µg

Lane 4:

human HEL whole cell lysates, at 30 µg

Lane 5:

rat brain tissue lysates, at 30 µg

Lane 6:

mouse brain tissue lysates. at 30 µg

Secondary

All lanes:

goat anti-rabbit IgG-HRP at 1/5000 dilution

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