Extracts prepared from 3T3L1 cells stimulated with LIF were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with STAT3 [pS727] antibody (Product # OPA1-03007) for 2 hours at room temperature in a 1% BSA-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphoserine-containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG HRP-conjugate (Product # ALI4404) and bands were detected using the Pierce SuperSignal™ method. The data show that only the peptide corresponding to STAT3 [pS727] blocks the antibody signal, demonstrating the specificity of the antibody.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide from human STAT3 containing Serine 727.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||Assay-Dependent|
* 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.
OPA1-03007 detects phosphorylated signal transducer and activator of transcription 3 (STAT3) from human and rat samples. This antibody is specific for phosphorylated STAT3 and does not cross-react with unphosphorylated STAT3. OPA1-03007 does not detect any other STATs or related proteins. Also reactive to rat STAT3.
OPA1-03007 has been successfully used in Western blot procedures.
OPA1-03007 immunogen is synthetic Peptide from human STAT3 containing Serine 727.
The signal transducer and activator of transcription (STAT) family of transcription factors is composed of seven family members (STATs 1, 2, 3, 4, 5A, 5B and 6) that are involved in many cell processes (apoptosis, anti-apoptosis, cell differentiation and proliferation) depending upon the cell type. STATs are activated by ligand binding of mostly cytokine receptors, that receptor association with a Janus kinase (JAK) family member that phosphorylates a STAT protein, causing it to translocate to the nucleus, where it can bind to DNA and regulate the transcription of particular genes. Phosphorylation on serine 727 by ERK1 and 2, JNK or MSK1, together with phosphorylation of tyrosine 705 by JAK1 and 2, is required for maximum transcriptional activity.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.