Anti-BDNF Antibody

筛选器： All IHC WB FCM

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  Figure 1. IHC analysis of BDNF using anti-BDNF antibody (PB9075).BDNF was detected in paraffin-embedded section of Rat 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-BDNF Antibody (PB9075) 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.

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  Figure 2. IHC analysis of BDNF using anti-BDNF antibody (PB9075).BDNF 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-BDNF Antibody (PB9075) 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.

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  Figure 3. Western blot analysis of BDNF using anti-BDNF antibody (PB9075). 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: Rat Brain Tissue LysateAfter 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 rabbit anti-BDNF antigen affinity purified polyclonal antibody (Catalog # PB9075) 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 1:10000 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 BDNF at approximately 38KD. The expected band size for BDNF is at 28KD.

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  Figure 4. Flow Cytometry analysis of U-87 cells using anti-BDNF antibody (PB9075).Overlay histogram showing U-87 cells stained with PB9075 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-BDNF Antibody (PB9075, 1:100) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 1:100) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1:100) used under the same conditions. Unlabelled sample (Red line) was also used as a control.