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HEK-Blue™ TGF-β Cells

HEK-Blue™ TGF-β cells Unit size Cat. code Docs Qty Price
3-7 x 10e6 cells
hkb-tgfb
+-
$1,142.00

TGF-β Sensor 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 HEK293 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™. TGF-β-mediated SEAP production can be blocked using a neutralizing antibody, such as anti-hTGFβ-IgA.


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.

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Specifications

Antibiotic resistance: blasticidin, hygromycin B, Zeocin™

Growth medium: DMEM, 4.5 g/l glucose, 2-4 mM L-glutamine, 10% (v/v) fetal bovine serum, 50 U/ml penicillin, 50 μg/ml streptomycin, 100 μg/ml Normocin™

Guaranteed mycoplasma-free

Shipped on dry ice

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

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Contents

• 1 vial containing 3-7 x 106 cells

• 100 μl Blasticidin (10 mg/ml)

• 100 μl Hygromycin B Gold (100 mg/ml)

• 100 μl Zeocin™ (100 mg/ml)

• 1 ml Normocin™ (50 mg/ml)

• 1 pouch of QUANTI-Blue™ (SEAP detection medium)

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Description

HEK-Blue TGF-beta pathway

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.

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Citations

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