Anti-Bim/BCL2L11 Antibody

• Catalog Number: orb1098118
• Category: Antibodies
• Description: Anti-Bim/BCL2L11 Antibody. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Mouse.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IHC, WB
• Reactivity: Human, Mouse
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human Bim/BCL2L11, which shares 80% amino acid (aa) sequence identity with mouse and rat BCL2L11.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 28 kDa
• UniProt ID: O43521
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Rho-related GTP-binding protein RhoB; Rho cDNA clo
• Note: For research use only
• Application notes: Western blot, 0.1-0.25 μg/ml, Human, Mouse Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of HL-60 cells using anti-Bim/BCL2L11 antibody. Overlay histogram showing HL-60 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Bim/BCL2L11 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IHC analysis of Bim/BCL2L11 using anti-Bim/BCL2L11 antibody. Bim/BCL2L11 was detected in a 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 2 µg/ml rabbit anti-Bim/BCL2L11 Antibody 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) with DAB as the chromogen.

IHC analysis of Bim/BCL2L11 using anti-Bim/BCL2L11 antibody. Bim/BCL2L11 was detected in a paraffin-embedded section of human lung 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 2 µg/ml rabbit anti-Bim/BCL2L11 Antibody 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) with DAB as the chromogen.

IHC analysis of Bim/BCL2L11 using anti-Bim/BCL2L11 antibody. Bim/BCL2L11 was detected in a paraffin-embedded section of human tonsil 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 2 µg/ml rabbit anti-Bim/BCL2L11 Antibody 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) with DAB as the chromogen.

Western blot analysis of Bim/BCL2L11 using anti-Bim/BCL2L11 antibody. 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 Raji whole cell lysates, Lane 2: human COLO-320 whole cell lysates, Lane 3: human SW620 whole cell lysates, Lane 4: human THP-1 whole cell lysates, Lane 5: mouse RAW264.7 whole cell lysates, Lane 6: mouse Ana-1 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-Bim/BCL2L11 antigen affinity purified polyclonal antibody at 0.25 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 with Tanon 5200 system. A specific band was detected for Bim/BCL2L11 at approximately 28 kDa. The expected band size for Bim/BCL2L11 is at 22 kDa.