Anti-CKAP4 Antibody Picoband®

Western blot analysis of CKAP4 using anti-CKAP4 antibody (A02154-2).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 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 Hela whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human U251 whole cell lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: rat RH35 whole cell lysates,
Lane 7: mouse NIH/3T3 whole cell lysates,
Lane 8: mouse HEPA1-6 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 rabbit anti-CKAP4 antigen affinity purified polyclonal antibody (A02154-2) at 1:10000 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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for CKAP4 at approximately 70 kDa. The expected band size for CKAP4 is at 66 kDa.

Western blot analysis of CKAP4 using anti-CKAP4 antibody (A02154-2).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hacat whole cell lysates,
Lane 2: human U251 whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human PC-3 whole cell lysates,
Lane 5: rat heart tissue lysates,
Lane 6: rat H9C2(2-1) whole cell lysates,
Lane 7: mouse heart tissue lysates,
Lane 8: mouse C2C12 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 rabbit anti-CKAP4 antigen affinity purified polyclonal antibody (A02154-2) 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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for CKAP4 at approximately 66 kDa. The expected band size for CKAP4 is at 66 kDa.

IHC analysis of CKAP4 using anti-CKAP4 antibody (A02154-2).
CKAP4 was detected in a paraffin-embedded section of human pancreas cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CKAP4 Antibody (A02154-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of CKAP4 using anti-CKAP4 antibody (A02154-2).
CKAP4 was detected in a paraffin-embedded section of human stomach cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-CKAP4 Antibody (A02154-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of CKAP4 using anti-CKAP4 antibody (A02154-2).
CKAP4 was detected in an immunocytochemical section of SiHa 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 rabbit anti-CKAP4 Antibody (A02154-2) overnight at 4°C. Fluoro488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 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.

Immunoprecipitating CKAP4 in Hacat whole cell lysate.
Western blot analysis of CKAP4 using anti-CKAP4 antibody (A02154-2).
Lane 1: Hacat whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-CKAP4 antibody in Hacat whole cell lysate,
Lane 3: anti-CKAP4 antibody (2μg) + Hacat whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-CKAP4 antigen affinity purified polyclonal antibody (A02154-2) at a dilution of 0.5 μg/mL and probed with a mouse anti-rabbit IgG-HRP secondary antibody (Catalog # BM2007). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for CKAP4 at approximately 66 kDa. The expected band size for CKAP4 is at 66 kDa.