Application Notes: Western blot, 0.25-0.5 µg/ml, Human Immunohistochemistry(Paraffin-embedded Section), 2-5 µg/ml, Human, Mouse Flow Cytometry (Fixed), 1-3 µg/1x106 cells, Human. Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml
Flow Cytometry analysis of PC-3 cells using anti-Poliovirus Receptor/PVR antibody. Overlay histogram showing PC-3 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Poliovirus Receptor/PVR Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-mouse IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
IHC analysis of Poliovirus Receptor/PVR using anti-Poliovirus Receptor/PVR antibody. Poliovirus Receptor/PVR was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-Poliovirus Receptor/PVR Antibody overnight at 4C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit with DAB as the chromogen.
IHC analysis of Poliovirus Receptor/PVR using anti-Poliovirus Receptor/PVR antibody. Poliovirus Receptor/PVR was detected in a paraffin-embedded section of mouse liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-Poliovirus Receptor/PVR Antibody overnight at 4C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit with DAB as the chromogen.
Western blot analysis of Poliovirus Receptor/PVR using anti-Poliovirus Receptor/PVR antibody. 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 ug of sample under reducing conditions. Lane 1: human A431 whole cell lysates, Lane 2: human A549 whole cell lysates, Lane 3: human HT1080 whole cell lysates, Lane 4: human Daudi whole cell lysates. 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 mouse anti-Poliovirus Receptor/PVR antigen affinity purified monoclonal antibody at 0.5 µg/mL overnight at 4C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-mouse 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 Poliovirus Receptor/PVR at approximately 70-80 kDa. The expected band size for Poliovirus Receptor/PVR is at 45 kDa.
IHC analysis of Poliovirus Receptor/PVR using anti-Poliovirus Receptor/
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