Figure 1. Western blot analysis of ALK-1/ACVRL1 using anti-ALK-1/ACVRL1 antibody (MBS1753435).
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 30ug of sample under reducing conditions.
Lane 1: human HL-60 whole cell lysates
Lane 2: human THP-1 whole cell lysates
Lane 3: rat brain tissue lysates
Lane 4: rat heart tissue lysates
Lane 5: mouse brain tissue lysates
Lane 6: mouse kidney tissue lysates
Lane 7: mouse heart tissue lysates.
After 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-ALK-1/ACVRL1 antigen affinity purified polyclonal antibody (Catalog # MBS1753435) at 0. 5 μg/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 (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for ALK-1/ACVRL1 at approximately 55KD. The expected band size for ALK-1/ACVRL1 is at 55KD.
Figure 2. IHC analysis of ALK-1/ACVRL1 using anti-ALK-1/ACVRL1 antibody (MBS1753435).
ALK-1/ACVRL1 was detected in paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-ALK-1/ACVRL1 Antibody (MBS1753435) 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) (Catalog # MBS176451) with DAB as the chromogen.
Figure 3. IHC analysis of ALK-1/ACVRL1 using anti-ALK-1/ACVRL1 antibody (MBS1753435).
ALK-1/ACVRL1 was detected in paraffin-embedded section of human gallbladder adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-ALK-1/ACVRL1 Antibody (MBS1753435) 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) (Catalog # MBS176451) with DAB as the chromogen.
Figure 4. IHC analysis of ALK-1/ACVRL1 using anti-ALK-1/ACVRL1 antibody (MBS1753435).
ALK-1/ACVRL1 was detected in paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-ALK-1/ACVRL1 Antibody (MBS1753435) 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) (Catalog # MBS176451) with DAB as the chromogen.
Figure 5. IHC analysis of ALK-1/ACVRL1 using anti-ALK-1/ACVRL1 antibody (MBS1753435).
ALK-1/ACVRL1 was detected in paraffin-embedded section of human ovarian serous adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-ALK-1/ACVRL1 Antibody (MBS1753435) 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) (Catalog # MBS176451) with DAB as the chromogen.
Figure 6. Flow Cytometry analysis of MCF-7 cells using anti-ALK-1/ACVRL1 antibody (MBS1753435).
Overlay histogram showing MCF-7 cells stained with MBS1753435 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ALK-1/ACVRL1 Antibody (MBS1753435, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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.
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