Bad Rabbit Monoclonal Antibody

• Catalog Number: orb548220
• Category: Antibodies
• Description: Anti-Bad Rabbit Monoclonal Antibody. Tested in WB, IHC, ICC/IF, IP, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.
• Clonality: Monoclonal
• Species/Host: Rabbit
• Isotype: Rabbit IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Form/Appearance: Liquid
• Concentration: 0.5mg/ml
• Purification: Affinity-chromatography
• Immunogen: A synthesized peptide derived from human Bad
• UniProt ID: Q92934
• MW: 23 kDa
• Tested applications: FC, ICC, IF, IHC, IP, WB
• Application notes: WB 1:500-2000IHC 1:50-200ICC/IF 1:50-200IP 1:30FC 1:50. Restore with deionized water (or equivalent) for reconstitution volume of 1.0 mL
• Antibody Type: Primary Antibody
• Clone Number: CHC-2
• 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: Bcl2-associated agonist of cell death; BAD; Bcl-2-
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Immunohistochemical analysis of paraffin-embedded human colon cancer, using Bad Antibody (orb548220) BAD was detected in paraffin-embedded tissue section. 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 1ug/ml rabbit anti-BAD Antibody (orb548220) 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 # orb90444) with DAB as the chromogen.

Western blot analysis of BAD expression in HeLa cell lysate (orb548220). 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. 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-BAD monoclonal antibody (Catalog # orb548220) 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 # orb90503) with Tanon 5200 system.

IF analysis of immunocytochemical section of HeLa cells using anti-Bad antibody (orb548220) Bad was detected in immunocytochemical section. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/mL rabbit anti-Bad Antibody (orb548220) overnight at 4 &deg.