TGF-β Reporter HEK 293 Cells
|HEK-Blue™ TGF-β cells||Unit size||Cat. code||Docs||Qty||Price|
Human TGFβ SEAP Reporter Cells
3-7 x 10e6 cells
TGF-β Reporter Cells
HEK-Blue™ TGF-β cells allow the detection of bioactive TGF-β by monitoring the activation of the TGF-β/Smad pathway.
They were generated by stable transfection of human embryonic kidney HEK 293 cells with the human TGFBRI, Smad3 and Smad4 genes. They further express a Smad3/4-binding elements (SBE)-inducible SEAP reporter gene.
Stimulation of HEK-Blue™ TGF-β cells with TGF-β induces the activation of the TGF-β/Smad signaling pathway leading to the formation of a Smad3/Smad4 complex.
The heterocomplex enters the nucleus and binds SBE sites inducing the production of SEAP. The quantity of SEAP secreted in the supernatant can be easily assessed using QUANTI-Blue™ Solution.
TGF-β-mediated SEAP production can be blocked using a neutralizing anti-hTGF-β antibody.
Features of HEK-Blue™ TGF-β cells:
- Fully functional TGF-β signaling pathway
- Readily assessable SEAP reporter activity
- Functionally tested and guaranteed mycoplasma-free
Applications of HEK-Blue™ TGF-β cells:
- Detection of human TGF-β
- Screening of anti-TGF-β antibodies
Stimulation of HEK-Blue™ TGF-β cells by recombinant human TGF-β was assessed by measuring the levels of SEAP using QUANTI-Blue™ and reading the optical density (O.D.) at 655 nm.
|TGF-β||0.2 +/- 0.1 ng/ml||22|
Note: The response ratio was calculated by dividing the OD at 655 nm for the treated cells by the OD at 655 nm for the untreated cells.
HEK-Blue™ TGF-β cells were stimulated with various human recombinant cytokines; IFNα (1000 IU/ml), IFNβ (1000 IU/ml), IFNγ (1000 IU/ml), IL-1β (100 ng/ml), IL-4 (100 ng/ml), IL-6 (100 ng/ml), IL-13 (100 ng/ml), IL-18 (100 ng/ml), TGF-β (10 ng/ml), TNF-α (100 ng/ml) and CD40L (100 ng/ml). After a 20 h incubation, SEAP activity was assessed using QUANTI-Blue™ and reading the O.D. at 655 nm.
Anti-hTGF-β-IgA was incubated with 1 ng/ml of hTGF-β for 4 h prior to the addition of HEK-Blue™ TGF-β cells, which were incubated with the antibody and cytokine for a further 16 h. Levels of SEAP in the supernatant were measured using QUANTI-Blue™ and reading the O.D. at 655 nm. The IC50 is 100 +/- 30 ng/ml.
Growth medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) heat-inactivated fetal bovine serum, 100 U/ml penicillin, 100 µg/ml streptomycin, 100 µg/ml Normocin™
Detects human TGF-β
Detection range for human TGF-β: 0.1 - 10 ng/mlBack to the top
- 1 vial containing 3-7 x 106 cells
- 1 ml of Blasticidin (10 mg/ml)
- 1 ml of Hygromycin B Gold (100 mg/ml)
- 1 ml of Zeocin™ (100 mg/ml)
- 1 ml of Normocin™ (50 mg/ml)
- 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent)
Shipped on dry ice (Europe, USA & Canada)Back to the top
Tumor growth factor-beta (TGF-β) belongs to a family of structurally related cytokines that regulate a plethora of cellular functions, such as proliferation, apoptosis, differentiation and migration.
TGF-β binds to a type II receptor which recruits and activates a type I receptor. The type I receptor then phosphorylates receptor-regulated Smads (R-Smads), such as Smad2 and Smad3, which associate with Smad4. R-Smad/Smad4 complexes accumulate in the nucleus where they regulate the transcription of target genes.
HEK-Blue™ TGF-β cells are designed to monitor TGF-β-induced Smad signaling, as these cells express a Smad3/4-binding elements (SBE)-inducible SEAP reporter gene.Back to the top