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Thermo Scientific™ LanthaScreen™ STAT5 TF-1 Cell Line
Description
LanthaScreen™ GFP Cellular Assays allow for the analysis of post-translational modifications for a number of target proteins in an live-cell context. In hematopoeitic cells, the JAK2/STAT5 signaling pathway plays an essential role in blood cell formation in response to cytokines such as GM-CSF, IL-3, and EPO. In this pathway, binding of these cytokines to their respective cell surface receptors results in the activation of JAK2, which in turn phospho-activates STAT5 proteins at specific tyrosine residues (Tyr-694/699).
LanthaScreen™-STAT5 TF-1 is a human hematopoeitic cell line which constitutively expresses GFP-STAT5 fusion proteins. The JAK2/STAT5 signaling pathway is known to be functionally intact in this cell line, therefore the GFP-STAT5 fusion protein serves as a substrate for the inducible phosphorylation by JAK2. Using this cell line, a homogenous immuno-assay has been developed in which the phosphorylation state of GFP-STAT5 is detected in cell lysates using a LanthaScreen™ Terbium-anti-mouse and anti-phospho STAT5 [pTyr694/699] antibody pair, in a time-resolved FRET (TR-FRET) readout. GFP-STAT5 Lentivirus was transduced into TF-1 cells followed by selection with Blasticidine.
This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker, and has been screened for the constitutive expression of GFP-STAT5 fusion protein. Using a lytic TR-FRET immuno-assay, this cell line is validated for EC50 and Z' under optimized conditions using GM-CSF as an agonist for JAK2 -mediated GFP-STAT5 phosphorylation. This assay has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and lysis/equilibration time.
Specifications
Specifications
| Alternate Agonists | IL-3, EPO |
| Antagonists | JAK Inhibitor |
| Assay Entry | Cell-based terbium phospho-specific activity (stable) |
| Cell State | Dividing Cells |
| Content And Storage | LanthaScreen™ STAT5 TF-1 cells are shipped on dry ice. Store in liquid nitrogen immediately upon receipt, or thaw for immediate use. |
| Detection Method | Fluorescence |
| Gene Symbol | STAT5A |
| Optimal Response | GM-CSF (EC50 = 0.33 ng/mL) |
| Primary Agonist | GM-CSF |
| Product Line | LanthaScreen™ |
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For Research Use Only. Not for use in diagnostic procedures.
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