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Western blot
ImmunohistochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
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- Product number
- A00237-3 - Provider product page
- Provider
- Boster Biological Technology
- Product name
- Anti-FADD Antibody Picoband™
- Antibody type
- Polyclonal
- Description
- Rabbit IgG polyclonal antibody for FADD detection. Tested with WB, IHC-P, FCM in Mouse;Rat.
- Reactivity
- Mouse, Rat
- Host
- Rabbit
- Vial size
- 100μg/vial
- Concentration
- Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
- Storage
- At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquoted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing.
- Handling
- Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
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Supportive validation
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- Boster Biological Technology (provider)
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- Experimental details
- Western blot analysis of FADD using anti-FADD antibody (A00237-3). 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 30ug of sample under reducing conditions. Lane 1: mouse brain tissue lysates, Lane 2: mouse stomach tissue lysates, Lane 3: mouse lung tissue lysates, Lane 4: mouse liver tissue lysates, Lane 5: mouse kidney tissue lysates, Lane 6: mouse spleen tissue lysates, Lane 7: mouse thymus tissue lysates, Lane 8: mouse NIH/3T3 whole cell lysates. 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-FADD antigen affinity purified polyclonal antibody (Catalog # A00237-3) 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 (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for FADD at approximately 30KD. The expected band size for FADD is at 23KD.
- Additional image
