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Dual Parameter Analysis of Total and Phospho STAT3 on A431Cells (A) Unstimulated A431 cells stained with total and phospho STAT1. 93% of the cells ar...
FlowCellect™ STAT3 Activation Dual Detection Kit
Description:
FlowCellect™ STAT3 Activation Dual Detection Kit
Trade Name:
FlowCellect
Qty/Pk:
25 Tests / Kit
Product Overview:
STAT3 is a transcription factor that is downstream of many signal transduction cascades that propagate signals generated extracellularly along intracellular interconnected transduction pathways. STAT3 pathway activation in response to receptor tyrosine kinase stimuli (e.g. EGFR stimulated by EGF) plays a significant role in the regulation of essential cellular processes. The diverse roles of STAT3 make it a key molecule in both normal and aberrant biology, as it can provide signals for cell survival, mitogenesis, as well as tumor-related stimuli for angiogenesis and metastasis.
Millipore’s FlowCellect™ STAT3 Activation Dual Detection kit includes two directly conjugated antibodies, a phospho-specific Anti-phospho-STAT3 (Tyr705)-Alexa Fluor® 647 and Anti-STAT3-Alexa Fluor® 488. The antibody pair provided in this kit has been carefully titrated to ensure the ability to measure Total and phospho-STAT3 simultaneously on the same protein for accurate determination of protein level and activation.
Sufficient reagents are provided to perform 25 two-color tests. Detailed assay instructions are included to assist in analysis and to ensure the correct cell concentration is obtained during acquisition of sample data.
Millipore’s FlowCellect™ STAT3 Activation Dual Detection kit includes two directly conjugated antibodies, a phospho-specific Anti-phospho-STAT3 (Tyr705)-Alexa Fluor® 647 and Anti-STAT3-Alexa Fluor® 488. The antibody pair provided in this kit has been carefully titrated to ensure the ability to measure Total and phospho-STAT3 simultaneously on the same protein for accurate determination of protein level and activation.
Sufficient reagents are provided to perform 25 two-color tests. Detailed assay instructions are included to assist in analysis and to ensure the correct cell concentration is obtained during acquisition of sample data.
Background Information:
Millipore’s FlowCellect™ Activation Dual Detection kits are a series of flow cytometry products which include a pair of antibodies that bind to the same protein; one to detect total protein expression and another to detect the phosphorylated form of the same target. Using two parameter analysis we can achieve target specific detection of phosphorylation and, by doing so, eliminate false positives while enhancing the signal to noise ratio
STATs are transcription factors that can be phosphorylated normally by a cell surface receptor such as cytokine receptors, followed by dimerization and nuclear translocation where they bind to DNA and activate transcription of their target genes. STAT3 (Signal transducer and activator of transcription 3) is a transcription factor that plays an important role in normal and aberrant mammalian biology. It becomes activated upon phosphorylation by receptor associated kinases such as JAK2, and responds to various types of upstream signaling pathways. Due to the diverse ways in which STAT3 can be activated in vivo, its biological functions vary, from regulating cell proliferation, to promoting oncogenesis, and even to act as a tumor suppressor in certain settings.
The regulation of STAT3 can be mediated through both protein expression modulation and post-translational modifications. The expression of STAT3 has been shown to be aberrantly elevated in disease states such as cervical cancer and some cases of large cell lymphoma, where abnormal cellular proliferation and survival were believed to be linked to rapid cancer development. In these cases, phosphorylation of STAT3 was concurrently observed with increased gene and protein expression, suggesting a system of combinatorial regulation of the protein and highlighting the importance of a dual-detection approach in studying cellular signaling pathways.
STATs are transcription factors that can be phosphorylated normally by a cell surface receptor such as cytokine receptors, followed by dimerization and nuclear translocation where they bind to DNA and activate transcription of their target genes. STAT3 (Signal transducer and activator of transcription 3) is a transcription factor that plays an important role in normal and aberrant mammalian biology. It becomes activated upon phosphorylation by receptor associated kinases such as JAK2, and responds to various types of upstream signaling pathways. Due to the diverse ways in which STAT3 can be activated in vivo, its biological functions vary, from regulating cell proliferation, to promoting oncogenesis, and even to act as a tumor suppressor in certain settings.
The regulation of STAT3 can be mediated through both protein expression modulation and post-translational modifications. The expression of STAT3 has been shown to be aberrantly elevated in disease states such as cervical cancer and some cases of large cell lymphoma, where abnormal cellular proliferation and survival were believed to be linked to rapid cancer development. In these cases, phosphorylation of STAT3 was concurrently observed with increased gene and protein expression, suggesting a system of combinatorial regulation of the protein and highlighting the importance of a dual-detection approach in studying cellular signaling pathways.
Key Applications:
Flow Cytometry
Species Reactivity:
Human
Usage Statement:
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
- Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Quality Assurance:
Lot to lot in-process testing on all kit components
Components:
- 1. 20X Anti-phospho-STAT3 (Tyr705) Alexa Fluor® 647: (Part No.CS206488) One vial containing 150 µL.
- 2. 20X Anti-STAT3-Alexa Fluor® 488: (Part No. CS206476) One vial containing 150 µL.
- 3. Fixation Buffer: (Part No. CS202122) One bottle containing 13 mL.
- 4. 10X Wash Buffer: (Part No. CS202123) One bottle containing 13 mL.
- 5. 5X Assay Buffer: (Part No. CS202124) One 55 mL bottle
- 6. 1X Permeabilization Buffer: (Part No. CS203284) One bottle containing 13 mL.
Host:
Mouse
Storage Conditions:
4-8°C for antibodies and buffers
Detection Methods:
Fluorescent
Product Name:
FlowCellect™ STAT3 Activation Dual Detection Kit
Alternate Names:
Flow Cellect
Flow Cytometry Software Type:
ExpressPro & inCyte
Antibody Type:
Monoclonal Antibody
Flow Cytometry Market:
Research