STAT5 Luciferase Reporter-Ba/F3 Cell Line

reporter cell line

Product Info Summary

SKU: RC1035
Size: 1 Vial, Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.
Application: Functional Assay

Product Name

STAT5 Luciferase Reporter-Ba/F3 Cell Line

SKU/Catalog Number

RC1035

Size

1 Vial, Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.

Description

The STAT5 Luciferase Reporter cell line is a stably transfected Ba/F3 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the STAT5 responsive promoter, so that the cell line is designed to measure the transcriptional activity of STAT5. As a transcription factor, Signal Transducer and Activator of Transcription 5 (STAT5) is activated through phosphorylation at tyrosine 694 in response to many cytokines and growth factors including IL-2, IL-3, GM-CSF and prolactin. Aberrant STAT5 activity is closely related to a wide range of human cancers as STAT5 is often found to be constitutively phosphorylated in cancer cells. The phosphorylated STAT5 forms homodimers or heterodimers with other STATs, and the dimers translocate to nucleus in which DNA binding/promoter induction occurs. The STAT5 induction by IL-3 is shown in Figure 1. 

Contents

Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.

Storage & Handling

Immediately upon receipt, store in liquid nitrogen.

Cite This Product

STAT5 Luciferase Reporter-Ba/F3 Cell Line (Boster Biological Technology, Pleasanton CA, USA, Catalog # RC1035)

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. Actual working concentration varies and should be decided by the user.

Application:

Monitor the STAT5 signaling pathway activity.Screen for activators or inhibitors of the STAT5 signaling pathway. 

Culture conditions:

 Cells should be grown at 37°C with 5% CO2 using RPMI medium supplemented with 10% FBS, 1 mM sodium pyruvate, 10 mM HEPES and 1% Pen/Strep, plus 1 ng/ml mIL-3 (Note: mIL-3 is essential for Ba/F3 cell maintenance), plus 3 µg/ml of Puromycin. It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37°C water-bath, transfer to a tube containing 10 ml of growth medium without Puromycin, spin down cells, resuspend cells in pre-warmed growth medium without Puromycin, transfer resuspended cells to T25 flask and culture in 37°C-CO2 incubator.  Monitor the cell viability by counting cells daily for 1~3 days until cells completely recover viability as cells are doubling daily. Once cells are over 90% confluent, harvest cells by centrifugation and passage cells. At first passage, switch to growth medium containing Puromycin. Cells should be split before they reach complete confluence. To passage the cells, transfer cells to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cell suspension into new culture vessels. Subcultivation ration = 1:10 to 1:20 weekly.

Functional validation:

A. Response of STAT5  Ba/F3 cells to mIL-3.1. Harvest STAT5 Ba/F3 cells and seed cells into a white solid-bottom 96-well microplate in 100 µl of growth medium without IL-3 at 1 x 10^5 cells/well. 

Validation Images & Assay Conditions

Gene/Protein Information For (Source: Uniprot.Org, NCBI)

Uniprot ID

Gene ID

Gene Name

Full Name

Weight

*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".

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