Anti-Cyclin D3/CCND3 Antibody

• Catalog Number: orb196273
• Category: Antibodies
• Description: Anti-Cyclin D3/CCND3 Antibody. Tested in WB applications. This antibody reacts with Human, Rat.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: WB
• Reactivity: Human, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human Cyclin D3 recombinant protein (Position: Q136-L292). Human Cyclin D3 shares 94% and 93% amino acid (aa) sequences identity with mouse and rat Cyclin D3, respectively.
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 33 kDa
• UniProt ID: P30281
• 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: G1/S-specific cyclin-D3; CCND3
• Note: For research use only
• Application notes: Western blot, 0.1-0.5μg/ml, Human, Rat. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

Western blot analysis of Cyclin D3 using anti-Cyclin D3 antibody. 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 Cyclin D3 protein 0.5 ng. 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-Cyclin D3 antigen affinity purified polyclonal antibody 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 1:10000 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 Cyclin D3 at approximately 25 KD. The expected band size for Cyclin D3 is at 25 KD.

Western blot analysis of Cyclin D4 using anti-Cyclin D4 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 50 ug of sample under reducing conditions. lane 1: rat testis tissue lysate, lane 2: rat thymus tissue lysate, lane 3: rat lung tissue lysate, lane 4: rat ovary tissue lysate, lane 5: JURKAT whole cell lysate, lane 6: A549 whole cell lysate, lane 7: MCF-7 whole cell lysate, lane 8: HELA whole cell lysate. 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-Cyclin D4 antigen affinity purified polyclonal antibody 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 1:10000 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 Cyclin D4 at approximately 33 KD. The expected band size for Cyclin D4 is at 33 KD.