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Western blot analysis of STAT3 was performed by loading 25ug of U2-OS lysate from STAT3 SMART pool siRNA (Product# L-003544-00-0005) transfected or non-targeting control (Product# D-001810-10) transfected U2-OS cells onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with STAT3 (Clone 9D8) mouse monoclonal antibody at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-mouse-HRP secondary antibody (Product # 32430) at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Pierce Super Signal West Dura (Product #34075).
|Tested species reactivity||Mouse|
|Host / Isotype||Goat / IgG|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with proprietary stabilizer|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:60-1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody may cross-react with immunoglobulins from other species.
Working dilutions for Western blotting using SuperSignal West Substrates:
SuperSignal West Pico Substrate - 1:250-1:1250
SuperSignal West Dura Substrate - 1:600-1:3000
SuperSignal West Femto Substrate - 1:1250-1:6000
ECL Western Blotting Substrate - 1:20-1:125
Thermo Scientific Anti-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for mouse immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.