Anti-PVRL1/NECTIN1 Antibody Picoband®

Figure 1. Western blot analysis of PVRL1/NECTIN1 using anti-PVRL1/NECTIN1 antibody (A04988-1).
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 Hek293 whole cell lysates,
Lane 2: human CCRF-CEM whole cell lysates,
Lane 3: human Sw620 whole cell lysates,
Lane 4: human U87 whole cell lysates,
Lane 5: human SGC-7901 whole cell lysates,
Lane 6: human Jurkat whole cell lysates,
Lane 7: rat PC-12 whole cell lysates,
Lane 8: mouse spleen tissue lysates,
Lane 9: mouse NIH/3T3 whole cell lysates,
Lane 10: mouse Raw264.7 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-PVRL1/NECTIN1 antigen affinity purified polyclonal antibody (Catalog # A04988-1) at 0.25 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 (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PVRL1/NECTIN1 at approximately 110KD. The expected band size for PVRL1/NECTIN1 is at 110KD.

Figure 2. IHC analysis of PVRL1/NECTIN1 using anti-PVRL1/NECTIN1 antibody (A04988-1).
PVRL1/NECTIN1 was detected in paraffin-embedded section of human rectal 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 1μg/ml rabbit anti-PVRL1/NECTIN1 Antibody (A04988-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Figure 3. IHC analysis of PVRL1/NECTIN1 using anti-PVRL1/NECTIN1 antibody (A04988-1).
PVRL1/NECTIN1 was detected in paraffin-embedded section of human rectal 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 1μg/ml rabbit anti-PVRL1/NECTIN1 Antibody (A04988-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Figure 4. IHC analysis of PVRL1/NECTIN1 using anti-PVRL1/NECTIN1 antibody (A04988-1).
PVRL1/NECTIN1 was detected in paraffin-embedded section of human rectal 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 1μg/ml rabbit anti-PVRL1/NECTIN1 Antibody (A04988-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Figure 5. IHC analysis of PVRL1/NECTIN1 using anti-PVRL1/NECTIN1 antibody (A04988-1).
PVRL1/NECTIN1 was detected in paraffin-embedded section of rat brain 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 1μg/ml rabbit anti-PVRL1/NECTIN1 Antibody (A04988-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Figure 6. IHC analysis of PVRL1/NECTIN1 using anti-PVRL1/NECTIN1 antibody (A04988-1).
PVRL1/NECTIN1 was detected in paraffin-embedded section of rat brain 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 1μg/ml rabbit anti-PVRL1/NECTIN1 Antibody (A04988-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Figure 7. Flow Cytometry analysis of PC-3 cells using anti-PVRL1/NECTIN1 antibody (A04988-1).
Overlay histogram showing PC-3 cells stained with A04988-1 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-PVRL1/NECTIN1 Antibody (A04988-1, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.