Flow Cytometry, Optimal dilutions should be determined by end users.
Flow Cytometry analysis of SiHa cells using anti-Poliovirus Receptor/PVR antibody (A00664-2). Overlay histogram showing SiHa cells stained with A00664-2 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Poliovirus Receptor/PVR Antibody (A00664-2, 1µg/1x106 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (BA1127, 5-10µg/1x106 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was rabbit IgG (1µg/1x106) used under the same conditions. Unlabelled sample without incubation w
IF analysis of Poliovirus Receptor/PVR using anti-Poliovirus Receptor/PVR antibody (A00664-2). Poliovirus Receptor/PVR was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5µg/mL rabbit anti-Poliovirus Receptor/PVR Antibody (A00664-2) overnight at 4C. DyLight488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
IHC analysis of Poliovirus Receptor/PVR using anti-Poliovirus Receptor/PVR antibody (A00664-2). Poliovirus Receptor/PVR was detected in 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 2µg/ml rabbit anti-Poliovirus Receptor/PVR Antibody (A00664-2) overnight at 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog SA1022) with DAB as the chromogen.
IHC analysis of Poliovirus Receptor/PVR using anti-Poliovirus Receptor/PVR antibody (A00664-2). Poliovirus Receptor/PVR was detected in paraffin-embedded section of human prostate 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 2µg/ml rabbit anti-Poliovirus Receptor/PVR Antibody (A00664-2) overnight at 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog SA1022) with DAB as the chromogen.
Western blot analysis of Poliovirus Receptor/PVR using anti-Poliovirus Receptor/PVR antibody (A00664-2). 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 30ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human A549 whole cell lysates, Lane 3: human HT1080 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Poliovirus Receptor/PVR antigen affinity purified polyclonal antibody (Catalog A00664-2) at 0.25 µg/mL overnight at 4C, 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 (Catalog EK1002) with Tanon 5200 system. A specific band was detected for Poliovirus Receptor/PVR at approximately 70-80KD. The expected band size for Poliovirus Receptor/PVR is at 70-80KD.
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