Anti-Calbindin/CALB1 Antibody Picoband®

Figure 1. Western blot analysis of Calbindin using anti-Calbindin antibody (A03047).
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: rat brain tissue lysates,
Lane 2: rat kidney tissue lysates,
Lane 3: mouse brain tissue lysates,
Lane 4: mouse kidney 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-Calbindin antigen affinity purified polyclonal antibody (Catalog # A03047) 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 Calbindin at approximately 30 kDa. The expected band size for Calbindin is at 30 kDa.

Figure 2. IHC analysis of Calbindin using anti-Calbindin antibody (A03047).
Calbindin was detected in paraffin-embedded section of rat kidney tissue. 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-Calbindin Antibody (A03047) 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 Calbindin using anti-Calbindin antibody (A03047).
Calbindin was detected in paraffin-embedded section of mouse brain tissue. 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-Calbindin Antibody (A03047) 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 Calbindin using anti-Calbindin antibody (A03047).
Calbindin was detected in paraffin-embedded section of mouse kidney tissue. 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-Calbindin Antibody (A03047) 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. Flow Cytometry analysis of A431 cells using anti-Calbindin antibody (A03047).
Overlay histogram showing A431 cells stained with A03047 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Calbindin Antibody (A03047,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight488 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 (Red line) was also used as a control.

Figure 6. IF analysis of Calbindin using anti-Calbindin antibody (A03047).
Calbindin was detected in a paraffin-embedded section of rat cerebellum 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 rabbit anti-Calbindin Antibody (A03047) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®488 Conjugated Avidin (BA1128). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 7. IF analysis of Calbindin using anti-Calbindin antibody (A03047).
Calbindin was detected in a paraffin-embedded section of rat cerebellum 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 rabbit anti-Calbindin Antibody (A03047) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using DyLight®488 Conjugated Avidin (BA1128). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.