Control Lysate
(HeLa cells were grown in a T175 flask at 37 degree C, 5%CO2, in DMEM with 10% FBS and cultured to 60% confluence. The medium was removed and cells were stimulated with 1 nM of hIL 4 for 20 min. The medium was removed and cells were lysed with 3 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.)
STAT6 assay kit
Phospho-STAT6 (Y641) + Total STAT6 TR-FRET Cellular Assay Kit
Predicted: Rat
Predicted: Rat
Acc-phospho-STAT5 (80 uL)
Eu-Total STAT5 (5 uL)
Acc-Total STAT5 (20 uL)
STAT5 control lysate (500 uL)
5X Lysis Buffer #2 (5 mL)
10X TR-FRET Detection buffer (250 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 60% confluence. The medium was removed and cells were stimulated with 1 nM of hIL 4 for 20 min. The medium was removed and cells were lysed with 3 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 60% confluence. The medium was removed and cells were stimulated with 1 nM of hIL 4 for 20 min. The medium was removed and cells were lysed with 3 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 hIL 4 for 20 min (stimulation) or JAK inhibitor 1for 30 min then 0.5 nM hIL 4 (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 on separate wells for phospho-STAT6 (Y641) and total-STAT6 using the two-plate assay protocol. 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 hIL 4 for 20 min (stimulation) or JAK inhibitor 1for 30 min then 0.5 nM hIL 4 (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 on separate wells for phospho-STAT6 (Y641) and total-STAT6 using the two-plate assay protocol. 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
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• TR-FRET technology :: Proven, sensitive and robust assay technology
• Homogeneous, add-and-read assay :: No wash, no separation steps
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NCBI and Uniprot Product Information
NCBI Description
The protein encoded by this gene is a member of the STAT family of transcription factors. 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 plays a central role in exerting IL4 mediated biological responses. It is found to induce the expression of BCL2L1/BCL-X(L), which is responsible for the anti-apoptotic activity of IL4. Knockout studies in mice suggested the roles of this gene in differentiation of T helper 2 (Th2) cells, expression of cell surface markers, and class switch of immunoglobulins. Alternative splicing results in multiple transcript variants.[provided by RefSeq, May 2010]
Uniprot Description
STAT6: transcription factor of the STAT family. Plays a central role in IL4-mediated biological responses. Induces the expression of BCL2L1/BCL-X(L), which is responsible for the anti-apoptotic activity of IL4. May function in the differentiation of T helper 2 cells and class switch of immunoglobulins. Forms homo- or heterodimers that translocate into the nucleus where they regulate transcription.
Protein type: DNA-binding; Transcription factor
Chromosomal Location of Human Ortholog: 12q13
Cellular Component: nucleoplasm; nuclear chromatin; cytoplasm; cytosol; lipid raft
Molecular Function: identical protein binding; signal transducer activity; protein binding; transcription factor activity; protein phosphatase binding
Biological Process: regulation of transcription from RNA polymerase II promoter; T-helper 1 cell lineage commitment; negative regulation of T-helper 2 type immune response; transcription, DNA-dependent; positive regulation of interferon type I production; innate immune response; mammary gland epithelial cell proliferation; positive regulation of transcription from RNA polymerase II promoter; negative regulation of transcription from RNA polymerase II promoter; signal transduction; positive regulation of isotype switching to IgE isotypes; regulation of cell proliferation
Research Articles on STAT6
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Product Notes
The Human, Mouse Predicted: Rat STAT6 stat6 (Catalog #AAA298067) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA298067 Assay Kit recognizes Human, Mouse Predicted: Rat STAT6. AAA Biotech's STAT6 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 STAT6 stat6 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 "STAT6, 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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