Control Lysate
(HeLa cells were grown in a T175 flask at 37 degree C, 5%CO2, in DMEM with 10% FBS and cultured to 90% confluence. The medium was removed and cells were stimulated with 3 nM of IFN-alpha for 30 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #2 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
STAT3 assay kit
Total STAT3 TR-FRET Cellular Assay Kit
Acc-Total STAT3 (20 uL)
STAT3 control lysate (100 uL)
5X Lysis Buffer #2 (1 mL)
10X TR-FRET Detection buffer (50 uL)
Control Lysate
(HeLa cells were grown in a T175 flask at 37 degree C, 5%CO2, in DMEM with 10% FBS and cultured to 90% confluence. The medium was removed and cells were stimulated with 3 nM of IFN-alpha for 30 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #2 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Control Lysate
(HeLa cells were grown in a T175 flask at 37 degree C, 5%CO2, in DMEM with 10% FBS and cultured to 90% confluence. The medium was removed and cells were stimulated with 3 nM of IFN-alpha for 30 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #2 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Testing Data
(HeLa cells were seeded at 40,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in DMEM with 10% FBS. Cells were treated with either IFN-alpha for 30 min (stimulation) or with JAK inhibitor 1for 30 min then 1.5 nM IFN-alpha (inhibition). The media was removed from the wells, and cells were lysed with 50 uL/well of 1X Lysis Buffer #2 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. The lysates were then analyzed for total-STAT3 using the two-plate assay protocol. Lysates analyzed on separate wells for phospho-STAT3 (Y705) are also shown for comparison. The TR-FRET signal was recorded at 665 and 615 nm (EnVision lamp excitation) after a 4 h incubation period at room temperature.)
Testing Data
(HeLa cells were seeded at 40,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in DMEM with 10% FBS. Cells were treated with either IFN-alpha for 30 min (stimulation) or with JAK inhibitor 1for 30 min then 1.5 nM IFN-alpha (inhibition). The media was removed from the wells, and cells were lysed with 50 uL/well of 1X Lysis Buffer #2 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. The lysates were then analyzed for total-STAT3 using the two-plate assay protocol. Lysates analyzed on separate wells for phospho-STAT3 (Y705) are also shown for comparison. The TR-FRET signal was recorded at 665 and 615 nm (EnVision lamp excitation) after a 4 h incubation period at room temperature.)
A superior alternative to ELISA
Features :: Benefits
• TR-FRET technology :: Proven, sensitive and robust assay technology
• Homogeneous, add-and-read assay :: No wash, no separation steps
• < 1 hour bench time :: Faster, easier than ELISA
• Adherent or suspension cells :: Higher flexibility
• Validated assays :: Results you can trust
• Small sample volume (15uL) :: Save precious samples
• Compatible with low-volume 96-well and 384-well plates : Higher throughput
• Signal stable over time :: Flexibility to read plate
• Kits available in 100 and 500 assay points :: Cost-effective and flexible
• Kits available for phospho, total and phospho+total protein :: Allows multiple protein measurements in one plate from one sample
NCBI and Uniprot Product Information
NCBI Description
The protein encoded by this gene is a member of the STAT protein family. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor associated kinases, and then form homo- or heterodimers that translocate to the cell nucleus where they act as transcription activators. This protein is activated through phosphorylation in response to various cytokines and growth factors including IFNs, EGF, IL5, IL6, HGF, LIF and BMP2. This protein mediates the expression of a variety of genes in response to cell stimuli, and thus plays a key role in many cellular processes such as cell growth and apoptosis. The small GTPase Rac1 has been shown to bind and regulate the activity of this protein. PIAS3 protein is a specific inhibitor of this protein. Mutations in this gene are associated with infantile-onset multisystem autoimmune disease and hyper-immunoglobulin E syndrome. Alternative splicing results in multiple transcript variants encoding distinct isoforms. [provided by RefSeq, Sep 2015]
Uniprot Description
STAT3: transcription factor of the STAT family. Phosphorylated and activated by receptor-associated kinases downstream of many cytokines and growth-factor receptors. Constitutively active in a number of human tumors. Forms homo- or heterodimers that translocate into the nucleus where they regulate transcription. Two alternatively spliced isoforms have been described.
Protein type: Transcription factor; Nuclear receptor co-regulator; Motility/polarity/chemotaxis; DNA-binding
Chromosomal Location of Human Ortholog: 17q21.31
Cellular Component: nucleoplasm; cytoplasm; mitochondrial inner membrane; plasma membrane; cytosol; nucleus
Molecular Function: protein dimerization activity; protein binding; ligand-dependent nuclear receptor activity; signal transducer activity; DNA binding; sequence-specific DNA binding; protein kinase binding; transcription factor binding; protein phosphatase binding; transcription factor activity; CCR5 chemokine receptor binding; glucocorticoid receptor binding
Biological Process: transcription from RNA polymerase II promoter; nerve growth factor receptor signaling pathway; viral reproduction; somatic stem cell maintenance; positive regulation of transcription, DNA-dependent; radial glial cell differentiation; thermoregulation; negative regulation of transcription from RNA polymerase II promoter; glucose homeostasis; signal transduction; response to estradiol stimulus; negative regulation of cell proliferation; astrocyte differentiation; regulation of transcription, DNA-dependent; protein import into nucleus; acute-phase response; negative regulation of glycolysis; positive regulation of Notch signaling pathway; response to drug; nervous system development; intracellular receptor-mediated signaling pathway; eating behavior; cytokine and chemokine mediated signaling pathway; regulation of multicellular organism growth; JAK-STAT cascade; cellular response to hormone stimulus; regulation of transcription from RNA polymerase II promoter; cell proliferation; response to ethanol; sexual reproduction; positive regulation of transcription from RNA polymerase II promoter; eye photoreceptor cell differentiation; cell motility; phosphorylation
Disease: Hyper-ige Recurrent Infection Syndrome, Autosomal Dominant; Autoimmune Disease, Multisystem, Infantile-onset
Research Articles on STAT3
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Product Notes
The Human, Mouse, Rat STAT3 stat3 (Catalog #AAA298062) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA298062 Assay Kit recognizes Human, Mouse, Rat STAT3. AAA Biotech's STAT3 can be used in a range of immunoassay formats including, but not limited to, Cell-Based Assays. Researchers should empirically determine the suitability of the STAT3 stat3 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "STAT3, Assay Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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