Anti-NKCC1/SLC12A2 Antibody Picoband™ (monoclonal, 6G7D2)

Figure 1. Western blot analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody (M03603).
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 A549 whole cell lysates,
Lane 2: human CACO-2 whole cell lysates,
Lane 3: human PC-3 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 mouse anti-NKCC1/SLC12A2 antigen affinity purified monoclonal antibody (Catalog # M03603) 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 NKCC1/SLC12A2 at approximately 200 kDa. The expected band size for NKCC1/SLC12A2 is at 131 kDa.

Figure 2. IHC analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody (M03603).
NKCC1/SLC12A2 was detected in a paraffin-embedded section of human colorectal adenocarcinoma 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 mouse anti-NKCC1/SLC12A2 Antibody (M03603) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.

Figure 3. IHC analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody (M03603).
NKCC1/SLC12A2 was detected in a paraffin-embedded section of mouse colon 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 mouse anti-NKCC1/SLC12A2 Antibody (M03603) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.

Figure 4. IHC analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody (M03603).
NKCC1/SLC12A2 was detected in a paraffin-embedded section of rat colon 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 mouse anti-NKCC1/SLC12A2 Antibody (M03603) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.

Figure 5. IF analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody (M03603).
NKCC1/SLC12A2 was detected in a paraffin-embedded section of human colorectal adenocarcinoma 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 5 μg/mL mouse anti-NKCC1/SLC12A2 Antibody (M03603) overnight at 4°C. Biotin conjugated goat anti-mouse IgG (BA1001) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®550 Conjugated Avidin (BA1134). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 6. IF analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody (M03603).
NKCC1/SLC12A2 was detected in a paraffin-embedded section of mouse colon 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 5 μg/mL mouse anti-NKCC1/SLC12A2 Antibody (M03603) overnight at 4°C. Biotin conjugated goat anti-mouse IgG (BA1001) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®550 Conjugated Avidin (BA1134). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 7. IF analysis of NKCC1/SLC12A2 using anti-NKCC1/SLC12A2 antibody (M03603).
NKCC1/SLC12A2 was detected in a paraffin-embedded section of rat colon 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 5 μg/mL mouse anti-NKCC1/SLC12A2 Antibody (M03603) overnight at 4°C. Biotin conjugated goat anti-mouse IgG (BA1001) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®550 Conjugated Avidin (BA1134). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.