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14-3-3 gamma/YWHAG Antibody
Description
Images & Validation
−| Tested Applications | ELISA, FC, WB |
|---|---|
| Reactivity | Human, Mouse, Rat |
| Application Notes |
Key Properties
−| Antibody Type | Primary Antibody |
|---|---|
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | Rabbit IgG |
| Immunogen | E.coli-derived human 14-3-3 gamma/YWHAG recombinant protein (Position: Y107-A138). |
| Molecular Weight | 28 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 |
| Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
| Disclaimer | For research use only |
Alternative Names
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Flow Cytometry analysis of U87 cells using anti-14-3-3 Gamma/YWHAG antibody. Overlay histogram showing U87 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-14-3-3 Gamma/YWHAG Antibody (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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of 14-3-3 Gamma/YWHAG using anti-14-3-3 Gamma/YWHAG 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 brain tissue lysates, Lane 2: mouse brain 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-14-3-3 Gamma/YWHAG 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 14-3-3 Gamma/YWHAG at approximately 28 KD. The expected band size for 14-3-3 Gamma/YWHAG is at 28 KD.
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14-3-3 gamma/YWHAG Antibody (orb654477)
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