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Catalog Number | orb692173 |
---|---|
Category | Antibodies |
Description | FADD Antibody |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | FC, IHC, WB |
Reactivity | Mouse, Rat |
Isotype | Rabbit IgG |
Immunogen | A synthetic peptide corresponding to a sequence of mouse FADD (AEKKNASVAGLVKALRTCRLNLVADLVEEAQES). |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Dilution range | Western blot, 0.1-0.25μg/ml, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse Flow Cytometry, 1-3μg/1x106 cells, Mouse |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 30 kDa |
UniProt ID | Q61160 |
Storage | Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles. |
Note | For research use only |
Application notes | Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Add 0.2ml of distilled water will yield a concentration of 500μg/ml. |
Expiration Date | 12 months from date of receipt. |
Western blot analysis of FADD using anti-FADD antibody (orb692173). 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 # orb692173) 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 # orb90503) with Tanon 5200 system. A specific band was detected for FADD at approximately 30KD. The expected band size for FADD is at 23KD.
Western blot analysis of FADD using anti-FADD antibody (orb692173). 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: rat brain tissue lysates, Lane 2: rat stomach tissue lysates, Lane 3: rat lung tissue lysates, Lane 4: rat liver tissue lysates, Lane 5: rat kidney tissue lysates, Lane 6: rat spleen tissue lysates, Lane 7: rat thymus tissue lysates, Lane 8: rat PC-12 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 # orb692173) 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 # orb90503) with Tanon 5200 system. A specific band was detected for FADD at approximately 30KD. The expected band size for FADD is at 23KD.
IHC analysis of FADD using anti-FADD antibody (orb692173). FADD was detected in a paraffin-embedded section of mouse liver tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-FADD Antibody (orb692173) 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.
Flow Cytometry analysis of HEPA1-6 cells using anti-FADD antibody (orb692173). Overlay histogram showing HEPA1-6 cells stained with orb692173 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-FADD Antibody (orb692173, 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 (Red line) was also used as a control.
WB | |
Bovine, Canine, Equine, Gallus, Porcine, Rabbit |
ELISA, FC, ICC, IF, IHC-P, WB | |
Human, Mouse, Rat | |
Rabbit | |
Polyclonal | |
Unconjugated |
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