Anti-GOLPH2/GOLM1 Antibody Picoband®

Figure 1. Western blot analysis of GOLPH2/GOLM1 using anti-GOLPH2/GOLM1 antibody (A02975-3).
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 U87 whole cell lysates,
Lane 2: human Caco-2 whole cell lysates,
Lane 3: rat testis tissue lysates,
Lane 4: rat brain tissue lysates,
Lane 5: mouse brain tissue lysates,
Lane 6: mouse Neuro-2a 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-GOLPH2/GOLM1 antigen affinity purified polyclonal antibody (Catalog # A02975-3) 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 GOLPH2/GOLM1 at approximately 73 kDa. The expected band size for GOLPH2/GOLM1 is at 73 kDa.

Figure 2. IF analysis of GOLPH2/GOLM1 using anti-GOLPH2/GOLM1 antibody (A02975-3).
GOLPH2/GOLM1 was detected in an 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 5 μg/mL rabbit anti-GOLPH2/GOLM1 Antibody (A02975-3) 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 3. Flow Cytometry analysis of Caco-2 cells using anti-GOLPH2/GOLM1 antibody (A02975-3).
Overlay histogram showing Caco-2 cells stained with A02975-3 (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-GOLPH2/GOLM1 Antibody (A02975-3, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/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 4. IHC analysis of GOLPH2/GOLM1 using anti-GOLPH2/GOLM1 antibody (A02975-3).
GOLPH2/GOLM1 was detected in a paraffin-embedded section of human esophageal squamous carcinoma 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-GOLPH2/GOLM1 Antibody (A02975-3) 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 GOLPH2/GOLM1 using anti-GOLPH2/GOLM1 antibody (A02975-3).
GOLPH2/GOLM1 was detected in a paraffin-embedded section of human lung 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-GOLPH2/GOLM1 Antibody (A02975-3) 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 GOLPH2/GOLM1 using anti-GOLPH2/GOLM1 antibody (A02975-3).
GOLPH2/GOLM1 was detected in a paraffin-embedded section of human cervical intraepithelial neoplasias 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-GOLPH2/GOLM1 Antibody (A02975-3) 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 GOLPH2/GOLM1 using anti-GOLPH2/GOLM1 antibody (A02975-3).
GOLPH2/GOLM1 was detected in a paraffin-embedded section of human placenta 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-GOLPH2/GOLM1 Antibody (A02975-3) 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.