Phospho-Tyrosine Rabbit Monoclonal Antibody

• Catalog Number: orb548666
• Category: Antibodies
• Description: Phospho-Tyrosine Rabbit Monoclonal Antibody
• Species/Host: Rabbit
• Clonality: Monoclonal
• Clone Number: AAOI-20
• Tested applications: FC, ICC, IF, IP, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: A synthesized peptide derived from Phosphotyrosine
• Concentration: Actual concentration vary by lot. Use suggested dilution ratio to decide dilution procedure.
• Dilution range: WB 1:500-1:2000ICC/IF 1:50-1:200IP 1:50FC 1:50
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• Storage: Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Western blot analysis of Phosphotyrosine expression in Jurkat cell lysate (orb548666). 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 50ug of sample under reducing conditions. 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-Tyrosine monoclonal antibody (Catalog # orb548666) overnight at 4 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 1:10000 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 Tyrosine.