Anti-ch TOG/CKAP5 Antibody Picoband® (monoclonal, 3C13)

Figure 1. Western blot analysis of Ch TOG/CKAP5 using anti-Ch TOG/CKAP5 antibody (M05324).
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 50ug of sample under reducing conditions.
Lane 1: human U-87MG whole cell lysates,
Lane 2: human COLO-320 whole cell lysates,
Lane 3: human SW620 whole cell lysates,
Lane 4: human HEPG2 whole cell lysates,
Lane 5: human CACO-2 whole cell lysates,
Lane 6: rat brain tissue lysates,
Lane 7: mouse brain tissue 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 mouse anti-Ch TOG/CKAP5 antigen affinity purified monoclonal antibody (Catalog # M05324) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for Ch TOG/CKAP5 at approximately 225KD. The expected band size for Ch TOG/CKAP5 is at 225KD.

Figure 2. IHC analysis of Ch TOG/CKAP5 using anti-Ch TOG/CKAP5 antibody (M05324).
Ch TOG/CKAP5 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 mouse anti-Ch TOG/CKAP5 Antibody (M05324) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

Figure 3. IHC analysis of Ch TOG/CKAP5 using anti-Ch TOG/CKAP5 antibody (M05324).
Ch TOG/CKAP5 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 2μg/ml mouse anti-Ch TOG/CKAP5 Antibody (M05324) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

Figure 4. IHC analysis of Ch TOG/CKAP5 using anti-Ch TOG/CKAP5 antibody (M05324).
Ch TOG/CKAP5 was detected in paraffin-embedded section of human renal 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 mouse anti-Ch TOG/CKAP5 Antibody (M05324) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

Figure 5. IHC analysis of Ch TOG/CKAP5 using anti-Ch TOG/CKAP5 antibody (M05324).
Ch TOG/CKAP5 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 2μg/ml mouse anti-Ch TOG/CKAP5 Antibody (M05324) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

Figure 6. Flow Cytometry analysis of A549 cells using anti-Ch TOG/CKAP5 antibody (M05324).
Overlay histogram showing A549 cells stained with M05324 (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- Ch TOG/CKAP5 Antibody (M05324, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse 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 7. IF analysis of Ch TOG/CKAP5 using anti-Ch TOG/CKAP5 antibody (M05324).
Ch TOG/CKAP5 was detected in an immunocytochemical section of Hela 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 5 μg/mL mouse anti-Ch TOG/CKAP5 Antibody (M05324) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) 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.