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SMAD5 Rabbit Polyclonal Antibody
Description
Research Area
Images & Validation
−| Tested Applications | WB |
|---|---|
| Dilution Range | Western blot, 0.25-0.5 μg/ml, Human, Monkey, Mouse, Rat |
| Reactivity | Human, Monkey, Mouse, Rat |
Key Properties
−| Antibody Type | Primary Antibody |
|---|---|
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | Rabbit IgG |
| Immunogen | A synthetic peptide corresponding to a sequence in the middle region of human SMAD5, which shares 95.5% amino acid (aa) sequence identity with both mouse and rat SMAD5. |
| Target | Mothers against decapentaplegic homolog 5 |
| Molecular Weight | 52 kDa |
| Purification | Immunogen affinity purified. |
| Conjugation | Unconjugated |
Storage & Handling
−| 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. |
|---|---|
| Form/Appearance | Lyophilized |
| Buffer/Preservatives | Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4. |
| Concentration | 500 µg/ml |
| Expiration Date | 12 months from date of receipt. |
| Disclaimer | For research use only |
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Western blot analysis of SMAD5 using anti-SMAD5 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 Hela whole cell lysates, Lane 2: human HepG2 whole cell lysates, Lane 3: human 293T whole cell lysates, Lane 4:monkey COS-7 whole cell lysates, Lane 5: human Caco-2 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-SMAD5 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 1:5000 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 SMAD5 at approximately 52-60 kDa. The expected band size for SMAD5 is at 52 kDa.

Western blot analysis of SMAD5 using anti-SMAD5 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: rat liver tissue lysates, Lane 2: mouse liver tissue 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-SMAD5 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 1:5000 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 SMAD5 at approximately 52-60 kDa. The expected band size for SMAD5 is at 52 kDa.
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SMAD5 Rabbit Polyclonal Antibody (orb654304)
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