Anti-MCL1 Antibody Picoband®

Western blot analysis of MCL1 using anti-MCL1 antibody (PB9132).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: recombinant human MCL1 protein 0.5 ng.
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-MCL1 antigen affinity purified polyclonal antibody (Catalog # PB9132) 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 MCL1 at approximately 40 kDa. The expected band size for MCL1 is at 40 kDa.

Western blot analysis of MCL1 using anti-MCL1 antibody (PB9132).
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 spleen tissue lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: human COLO320 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-MCL1 antigen affinity purified polyclonal antibody (Catalog # PB9132) 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 MCL1 at approximately 37 kDa. The expected band size for MCL1 is at 37 kDa.

CYD0281 promotes HUVECs cell apoptosis by the exposure of the Bcl-2 BH3 domain. A Cell viability of HUVECs at 48 h. The exposure of the BH3 domain of Bcl-2 detected by immunofluorescence (IF) staining using anti-Bcl-2/BH3 domain antibody ( B ), and flow cytometry analysis of cell apoptosis ( C ) in HUVECs treated with BDA-366 and CYD0281 at the IC 50 concentration. HUVECs were treated with CYD0281 at the IC 50 concentration for 48 h, mitochondrial dysfunction was analyzed by Mito-Tracker Red CMXRos staining ( D ), Cyt c release ( E ) and PARP cleavage ( F ) were analyzed by western blotting assay, and Bcl-2 associated Bax or Bim was analyzed by co-IP using Bcl-2 antibody ( G ). H The expression of Bcl-2 in HUVECs was inhibited using siRNAs (1, 2, and 3) and analyzed by western blotting assay. I Flow cytometry analysis of cell apoptosis in Bcl-2-silenced HUVECs that were treated with CYD0281 ( n = 3). Significant effect compared to the DMSO group: * P < 0.05 and *** P < 0.001. ns: no significant differences. Scale bars: 50 μm
Index in PubMed under a CC BY license. PMID: 37237269

CYD0281 inhibits angiogenic differentiation of HUVECs in vitro. Representative microscopic images and quantification of cell migration ( A ), and tube formation in Matrigel ( B ). C Representative images of endothelial cells sprout and quantification from rat aortic ring assay at 8 days of treatment. Significant effect compared to the DMSO group: ** P < 0.01 and *** P < 0.001. Scale bars: 100 μm ( A and B ) and 500 μm ( C )
Index in PubMed under a CC BY license. PMID: 37237269

IHC analysis of MCL1 using anti-MCL1 antibody (PB9132).
MCL1 was detected in a paraffin-embedded section of rat intestine 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 1 μg/ml rabbit anti-MCL1 Antibody (PB9132) 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.

IHC analysis of MCL1 using anti-MCL1 antibody (PB9132).
MCL1 was detected in a paraffin-embedded section of human intestinal 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 1 μg/ml rabbit anti-MCL1 Antibody (PB9132) 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.

IHC analysis of MCL1 using anti-MCL1 antibody (PB9132).
MCL1 was detected in a paraffin-embedded section of mouse intestine 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 1 μg/ml rabbit anti-MCL1 Antibody (PB9132) 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.