Thermo Scientific™

LanthaScreen™ STAT5 TF-1 Cell Line

LanthaScreen™ GFP Cellular Assays allow for the analysis of post-translational modifications for a number of target proteins in an live-cell자세히 알아보기

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.

사양

대체 작용제IL-3, EPO

길항제JAK Inhibitor

어세이 입력Cell-based terbium phospho-specific activity (stable)

셀 상태Dividing Cells

검출 방법Fluorescence

유전자 기호STAT5A

최적 반응GM-CSF (EC50 = 0.33 ng/mL)

일차 작용제GM-CSF

제품라인LanthaScreen™

수량1 Vial

판독TR-FRET

리포터 유전자GFP (EmGFP)

타겟 입력STAT5

유형Cell Line

세포주TF-1

System TypeLanthaScreen™

Druggable TargetSignaling Pathway

Unit SizeEach