Anti-CRM1/XPO1 Antibody Picoband®

Figure 1. Western blot analysis of CRM1 using anti-CRM1 antibody (PB9966).
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 Hela whole cell lysates,
Lane 2: human COLO-320 whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: human A549 whole cell lysates,
Lane 5: human A431 whole cell lysates,
Lane 6: human SGC-7901 whole cell lysates,
Lane 7: human 22RV1 whole cell lysates,
Lane 8: rat heart tissue lysates,
Lane 9: rat skeletal muscle tissue lysates,
Lane 10: rat lung tissue lysates,
Lane 11: rat testis tissue lysates,
Lane 12: mouse heart tissue lysates,
Lane 13: mouse skeletal muscle tissue lysates,
Lane 14: mouse lung tissue lysates,
Lane 15: mouse testis tissue lysates,
Lane 16: mouse ovary tissue 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 rabbit anti-CRM1 antigen affinity purified polyclonal antibody (Catalog # PB9966) 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 (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CRM1 at approximately 123 kDa. The expected band size for CRM1 is at 123 kDa.

Figure 2. IHC analysis of CRM1 using anti-CRM1 antibody (PB9966).
CRM1 was detected in paraffin-embedded section of mouse intestine tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CRM1 Antibody (PB9966) 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 CRM1 using anti-CRM1 antibody (PB9966).
CRM1 was detected in paraffin-embedded section of rat testis tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CRM1 Antibody (PB9966) 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 CRM1 using anti-CRM1 antibody (PB9966).
CRM1 was detected in paraffin-embedded section of human intestinal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CRM1 Antibody (PB9966) 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 CRM1 using anti-CRM1 antibody (PB9966).
CRM1 was detected in frozen section of mouse intestine tissue . The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CRM1 Antibody (PB9966) 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 CRM1 using anti-CRM1 antibody (PB9966).
CRM1 was detected in frozen section of rat intestine tissue . The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CRM1 Antibody (PB9966) 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. IHC analysis of CRM1 using anti-CRM1 antibody (PB9966).
CRM1 was detected in frozen section of rat testis tissue . The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CRM1 Antibody (PB9966) 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 8. IF analysis of CRM1 using anti-CRM1 antibody (PB9966).
CRM1 was detected in immunocytochemical section of U20S cell. 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 2μg/mL rabbit anti-CRM1 Antibody (PB9966) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 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.

Figure 9. Flow Cytometry analysis of SiHa cells using anti-CRM1 antibody (PB9966).
Overlay histogram showing SiHa cells stained with PB9966 (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 rabbit anti-CRM1 Antibody (PB9966,1μg/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.

Figure 10. Flow Cytometry analysis of U87 cells using anti-CRM1 antibody (PB9966).
Overlay histogram showing U87 cells stained with PB9966 (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 rabbit anti-CRM1 Antibody (PB9966,1μg/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.