Anti-Alkaline Phosphatase, Tissue Non-Specific 抗体

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody (AB65834)

ALPL was detected in an immunocytochemical section of U2OS 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 5 µg/mL rabbit anti-ALPL Antibody (ab65834) 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. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody (AB65834)

ALPL was detected in a paraffin-embedded section of human acinic cell carcinoma 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 2 µg/ml rabbit anti-ALPL Antibody (ab65834) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody (AB65834)

Immunohistochemistry analysis of paraffin-embedded sections of human parotid acinar cell carcinoma tissue staining ALPL with ab65834 at 2 µg/ml. 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 the primary antibody overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and 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-Alkaline Phosphatase, Tissue Non-Specific antibody (AB65834)

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 ab65834 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 ALPL at approximately 80 kDa. The expected band size for ALPL is at 57 kDa.

All lanes:

Western blot - Anti-Alkaline Phosphatase, Tissue Non-Specific antibody (ab65834) at 0.5 µg/mL

All lanes:

human Hela whole cell lysates at 30 µg

Secondary

All lanes:

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

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