Figure 1. Western blot analysis of 5HT1B/HTR1B using anti-5HT1B/HTR1B antibody (MBS1754993).
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 A431 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: rat brain tissue lysates,
Lane 4: rat C6 whole cell lysates,
Lane 5: mouse brain tissue lysates,
Lane 7: 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-5HT1B/HTR1B antigen affinity purified polyclonal antibody (#A04887-1) at 0.5 ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for 5HT1B/HTR1B at approximately 50 kDa. The expected band size for 5HT1B/HTR1B is at 50 kDa.
Figure 2. IHC analysis of 5HT1B/HTR1B using anti-5HT1B/HTR1B antibody (MBS1754993).
5HT1B/HTR1B was detected in a paraffin-embedded section of human pancreatic 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 ug/ml rabbit anti-5HT1B/HTR1B Antibody (MBS1754993) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Figure 3. IHC analysis of 5HT1B/HTR1B using anti-5HT1B/HTR1B antibody (MBS1754993).
5HT1B/HTR1B was detected in a paraffin-embedded section of human rectal 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 ug/ml rabbit anti-5HT1B/HTR1B Antibody (MBS1754993) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Figure 4. IHC analysis of 5HT1B/HTR1B using anti-5HT1B/HTR1B antibody (MBS1754993).
5HT1B/HTR1B was detected in a paraffin-embedded section of mouse brain 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 ug/ml rabbit anti-5HT1B/HTR1B Antibody (MBS1754993) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Figure 5. IHC analysis of 5HT1B/HTR1B using anti-5HT1B/HTR1B antibody (MBS1754993).
5HT1B/HTR1B was detected in a paraffin-embedded section of rat brain 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 ug/ml rabbit anti-5HT1B/HTR1B Antibody (MBS1754993) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Figure 6. Flow Cytometry analysis of A549 cells using anti-5HT1B/HTR1B antibody (MBS1754993).
Overlay histogram showing A549 cells stained with MBS1754993 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-5HT1B/HTR1B Antibody (MBS1754993, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG (MBS177405, 5-10 ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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