Anti-14-3-3 zeta/delta/YWHAZ Antibody

• Catalog Number: orb413035
• Category: Antibodies
• Description: Anti-14-3-3 zeta/delta/YWHAZ Antibody. Tested in IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ICC, IF, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence in the middle region of human 14-3-3 zeta/delta, which shares 97.8% amino acid (aa) sequence identity with both mouse and rat 14-3-3 zeta/delta.
• 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: 28 kDa
• UniProt ID: P63104
• 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: 14-3-3 protein zeta/delta; Protein kinase C inhibi
• Note: For research use only
• Application notes: Western blot, 0.1-0.5μg/ml Immunocytochemistry/Immunofluorescence, 5μg/ml. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

IF analysis of 14-3-3 zeta/delta using anti-14-3-3 zeta/delta antibody. 14-3-3 zeta/delta was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL rabbit anti-14-3-3 zeta/delta Antibody overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of 14-3-3 zeta/delta using anti-14-3-3 zeta/delta 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 HepG2 whole cell lysates, Lane 2: human THP-1 whole cell lysates, Lane 3: human Raji whole cell lysates, Lane 4: human A549 whole cell lysates, Lane 5: human A431 whole cell lysates, Lane 6: human PC-3 whole cell lysates, Lane 7: human MCF-7 whole cell lysates, Lane 8: human SiHa 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-14-3-3 zeta/delta 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 zeta/delta at approximately 28 kDa. The expected band size for 14-3-3 zeta/delta is at 28 kDa.

Western blot analysis of 14-3-3 zeta/delta using anti-14-3-3 zeta/delta 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 brain tissue lysates, Lane 2: rat kidney tissue lysates, Lane 3: rat RH35 whole cell lysates, Lane 4: rat PC-12 whole cell lysates, Lane 5: mouse brain tissue lysates, Lane 6: mouse kidney tissue lysates, Lane 7: mouse ANA-1 whole cell lysates, Lane 8: mouse Neuro-2a 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-14-3-3 zeta/delta 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 zeta/delta at approximately 28 kDa. The expected band size for 14-3-3 zeta/delta is at 28 kDa.