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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
hkb-tgfb
+-
$1,260.00

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TGF-β Reporter Cells

HEK-Blue™ TGF-β Cells signaling pathway
HEK-Blue™ TGF-β Cells signaling pathway

HEK-Blue™ TGF-β cells allow the detection of bioactive transforming growth factor-beta (TGF-β) by monitoring the activation of the TGF-β/Smad pathway. TGF-β, also known as tumor growth factor-beta, is a secreted cytokine that regulates numerous cellular functions, such as cell proliferation, apoptosis, differentiation and migration [1, 2].

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Cell line description:

HEK-Blue™ TGF-β cells 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

 

References:

1.  Travis MA. & Sheppard D., 2014. TGF-β activation and function in immunity. Annu Rev Immunol. 32:51-82.
2. Taylor AW., 2009. Review of the activation of TGF-beta in immunity. J Leukoc Biol. 85(1):29-33.

Figures


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.

Ligand EC50 Response Ratio
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.

Neutralization of TGF-β  response
Neutralization of TGF-β  response

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.

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Specifications

Antibiotic resistance: blasticidin, hygromycin B, Zeocin™

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™

Guaranteed mycoplasma-free

Detects human TGF-β

Detection range for human TGF-β: 0.1 - 10 ng/ml

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Contents

Shipped on dry ice (Europe, USA & Canada)

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Details

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 [1,2]. TGF-β exists in at least three isoforms; TGF-β1, TGF-β2, and TGF-β3. In the immune system, TGF-β1 is the predominant isoform [1]. It is produced by many cell types, including macrophages, in a latent form that is bound to two other polypeptides, latent TGF-β1 binding protein (LTBP) and latency-associated peptide (LAP).

Upon cleavage of these proproteins, the mature TGF-β1 is released. This mature protein can bind its cell surface receptors and initiate signaling. 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.

 

1.  Travis MA. & Sheppard D., 2014. TGF-β activation and function in immunity. Annu Rev Immunol. 32:51-82.
2. Taylor AW., 2009. Review of the activation of TGF-beta in immunity. J Leukoc Biol. 85(1):29-33.

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FAQ

Visit our FAQ Any questions about our cell lines ? Visit our frequently asked questions page

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Citations

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