Recombinant human TGF-β1 protein - Bioactive cytokine

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Human TGF-β1 - Mammalian cell-expressed, tag-free, with HSA

Recombinant TGF-β1 is a high-quality and biologically active cytokine, validated using proprietary TGF-β reporter cells. This TGF-β isoform is produced in HEK293 cells to ensure protein glycosylation and bona fide 3D structure.

Recombinant human TGF-β1 can be used together with HEK-Blue™ TGF-β cells for the screening of inhibitory molecules, such as Fresolimumab, a monoclonal antibody targeting all isoforms of TGF-β (see figures).

TGF-β signaling and biological functions

Key features

• Each lot is validated using HEK-Blue™ TGF-β cells
• Endotoxin ≤ 0.1 EU/µg
• 0.2 µm sterile-filtered

Applications

• TGF-β detection and quantification assays (positive control)
• Screening and release assays for antibodies blocking TGF-β signaling
• Screening and release assays for engineered TGF-β proteins

Transforming growth factor-beta (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.

 More details

All InvivoGen products are for internal research use only, and not for human or veterinary use.

Figures

SDS PAGE of the recombinant human (h)TGFβ1 protein. 1.5 μg of hTGF-β1, reduced or non-reduced, was loaded on a 12% Mini-PROTEAN® TGX Stain-Free™ Precast Gel (Bio-Rad). Detection was performed as per the manufacturer’s instructions. The bands were detected at ~15 kDa (reduced condition) and ~25 kDa (non-reduced condition).

Dose-response of HEK-Blue™ TGF-β cells to recombinant TGF-β. Cells were stimulated with increasing concentrations of recombinant human (h)TGF-β. After overnight incubation, the Smad response was determined using QUANTI‑Blue™ Solution, a SEAP detection reagent, and reading the optical density (OD) at 650 nm. Data are shown as mean ± SEM.

Dose-dependent inhibition of HEK‑Blue™ TGF-β cellular response using Fresolimumab biosimilar. Increasing concentrations of Anti-hTGF-β-hIgG4 (S228P) (0.1 ng/ml - 10 µg/ml) were incubated with recombinant hTGF-β(3 ng/ml) for 1 hour before the addition of HEK‑Blue™ TGF-β cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown as the percentage of activity (mean ± SEM).

Specifications

Source: HEK293 cells

Species: Human

Alternative name: Transforming growth factor beta-1

Carrier: HSA (human serum albumin)

Tag: Tag-free

Accession number: P01137

Molecular weight: ~ 25 kDa (SDS PAGE, non-reducing conditions)

Solubility: 100 μg/ml in water

Formulation: Phosphate buffer saline (pH 7.4), 5% saccharose, 2% HSA

Sterility: 0.2 µm filtration

Form:  Lyophilized

Reconstitution buffer:  Endotoxin-free water (provided)

Purity:  ≥95% (SDS-PAGE)

Endotoxin:  The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK‑Blue™ TLR4 cells.

Tested applications:  Cellular assays

Quality control: Each lot is functionally tested and validated

Contents

Recombinant human TGF-β1 is provided as a lyophilized powder and is available in two quantities:

• rcyc-htgfb1: 10 μg
• rcyc-htgfb1-5: 50 μg (5 x 10 μg)
• 1.5 ml endotoxin-free water for rcyc-htgfb1 and rcyc-htgfb1-5

 Recombinant human TGF-β1 is shipped at room temperature.

Upon receipt, the product should be stored at -20°C.

 Avoid repeated freeze-thaw cycles.

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 type II receptors (TβR-II) which recruit and activate type I receptors (TβR-I). The active ligand-heterotetrameric receptor complex signals through downstream transcriptional factors named Smads, including Smad2, Smad3, and Smad4.
Smad complexes translocate into the nucleus where they regulate the transcription of target genes, which contain one or more Smad binding elements (SBEs) in their promoter region [3]. Perturbations in TGF-β signaling affect immune homeostasis and tolerance, leading to inflammatory diseases and tumor immune evasion [3].

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.
3. Battle E. & Massagué J., 2019. Transforming Growth Factor-beta Signaling in Immunity and Cancer. Immunity. 50(4):924.