Control Lysate
(A431 cells were grown in a T175 flask at 37 degree C, 5%CO2, in DMEM with 10% FBS and cultured to 95% confluence. The medium was removed and cells were stimulated with 300 nM of EGF for 10 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #4 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.)
STAT5 assay kit
Phospho-STAT5 (Y694/Y699) + Total STAT5 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 #4 (5 mL)
10X TR-FRET Detection buffer (250 uL)
Control Lysate (MBS298098)
Control Lysate
(A431 cells were grown in a T175 flask at 37 degree C, 5%CO2, in DMEM with 10% FBS and cultured to 95% confluence. The medium was removed and cells were stimulated with 300 nM of EGF for 10 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #4 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
(A431 cells were grown in a T175 flask at 37 degree C, 5%CO2, in DMEM with 10% FBS and cultured to 95% confluence. The medium was removed and cells were stimulated with 300 nM of EGF for 10 min. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #4 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
(A431 cells were seeded at 75,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in DMEM with 10% FBS. Cells were treated with either EGF for 10 min (stimulation) or erlotinibfor 15 min then 70 nM EGF (inhibition). The media was removed from the wells, and cells were lysed with 25 uL/well of 1X Lysis Buffer #4 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-STAT5 (Y694/Y699) and total-STAT5 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
(A431 cells were seeded at 75,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in DMEM with 10% FBS. Cells were treated with either EGF for 10 min (stimulation) or erlotinibfor 15 min then 70 nM EGF (inhibition). The media was removed from the wells, and cells were lysed with 25 uL/well of 1X Lysis Buffer #4 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-STAT5 (Y694/Y699) and total-STAT5 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
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 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 is activated by, and mediates the responses of many cell ligands, such as IL2, IL3, IL7 GM-CSF, erythropoietin, thrombopoietin, and different growth hormones. Activation of this protein in myeloma and lymphoma associated with a TEL/JAK2 gene fusion is independent of cell stimulus and has been shown to be essential for tumorigenesis. The mouse counterpart of this gene is found to induce the expression of BCL2L1/BCL-X(L), which suggests the antiapoptotic function of this gene in cells. Alternatively spliced transcript variants have been found for this gene. [provided by RefSeq, Dec 2013]
Uniprot Description
STAT5A: transcription factor of the STAT family. Phosphorylated and activated by receptor-associated kinases downstream of many cytokines and growth-factor receptors. Activation of this protein in myeloma and lymphoma associated with a TEL/JAK2 fusion protein is essential for the tumorigenesis. Induces the expression of BCL2L1/BCL-X(L) in the mouse, suggesting an antiapoptotic function of this protein. Forms homo- or heterodimers that translocate into the nucleus where they regulate transcription. Two alternatively spliced isoforms have been described.
Protein type: Motility/polarity/chemotaxis; Oncoprotein; Transcription factor; DNA-binding
Chromosomal Location of Human Ortholog: 17q11.2
Cellular Component: nucleoplasm; cytosol
Molecular Function: signal transducer activity; protein binding; transcription factor activity
Biological Process: lactation; succinate metabolic process; peptidyl-tyrosine phosphorylation; oxaloacetate metabolic process; positive regulation of interleukin-2 biosynthetic process; T cell differentiation in the thymus; female pregnancy; positive regulation of multicellular organism growth; fatty acid metabolic process; positive regulation of mitotic cell cycle; positive regulation of activated T cell proliferation; 2-oxoglutarate metabolic process; positive regulation of natural killer cell differentiation; sequestering of lipid; negative regulation of mast cell apoptosis; natural killer cell differentiation; allantoin metabolic process; luteinization; development of secondary male sexual characteristics; regulation of steroid metabolic process; creatinine metabolic process; T cell homeostasis; positive regulation of gamma-delta T cell differentiation; Peyer's patch development; isoleucine metabolic process; negative regulation of erythrocyte differentiation; transcription, DNA-dependent; valine metabolic process; citrate metabolic process; regulation of multicellular organism growth; development of secondary female sexual characteristics; JAK-STAT cascade; positive regulation of natural killer cell mediated cytotoxicity; creatine metabolic process; regulation of transcription from RNA polymerase II promoter; regulation of epithelial cell differentiation; positive regulation of B cell differentiation; positive regulation of transcription from RNA polymerase II promoter; taurine metabolic process; positive regulation of inflammatory response
Research Articles on STAT5
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Product Notes
The Human, Mouse Predicted: Rat STAT5 stat5a (Catalog #AAA298064) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA298064 Assay Kit recognizes Human, Mouse Predicted: Rat STAT5. AAA Biotech's STAT5 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 STAT5 stat5a 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 "STAT5, 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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