Phospho-Stat3 (Y705)

Cat Number: MAB-94616
Conjugate: Unconjugated
Size: 100 ug
Clone: EP21472
Concentration: 1mg/ml
Host: Rb
Isotype: IgG
Immunogen: A synthesized peptide derived from human STAT3
Reactivity: Hu, Ms, Rt

Western Blot: 1:1000 Immunohistochemistry: 1:200 Immunocytochemistry: 1:100 Immunofluorescence: 1:100 Immunoprecipitation: 1-2,5ug/mg lysate

Purification: Affinity-chromatography
Form: liquid
Buffer: Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.
Storage: Store at –20°C. Do not aliquot the antibody.

Western blot analysis of Phospho-STAT3 (Tyr705) expression in (1) HeLa cell lysate; (2) HeLa cell lysate treated with IFN-a. 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- STAT3 monoclonal antibody 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 1:10000 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 STAT3


Immunohistochemical analysis of paraffin-embedded human pancreas, using Phospho-STAT3 (Y705) Antibody STAT3 was detected in paraffin-embedded tissue section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-STAT3 Antibody overnight at 4°C. Biotinylated goat antirabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)with DAB as the chromogen.