Tgf β1 human elisa kit

The concentrations of CXCL13 and TGF-β1 in the supernatant were measured with a Human CXCL13/BLC/BCA-1 Quantikine ELISA Kit (R&D Systems) and TGF-β1 Human ELISA Kit (Thermo Fisher Scientific) according to the manufacturer’s instructions.

The ability of a-TGFβ (1D11) and nonspecific IgG-TGFβRII (anti-gp120-TGFβRII) antibody to equally bind TGFβ in vitro was evaluated by a standard ELISA assay (Supplementary Figure 1). rhTGFβ1 (0–2000 pg ml⁻¹) was added to the plates coated with either TGFβRII-Fc (R&D Systems), a-TGFβ (Bioxcel), or IgG-TGFβRII (1 μg ml⁻¹ each), and binding to rhTGFβ1 was detected by a biotinylated anti-human TGFβ1 antibody (R&D Systems). TGFβRII-Fc-coated plates were used as a TGFβ-binding positive control (Supplementary Figure 1). To demonstrate that both agents were administered at doses sufficient to saturate systemic TGFβ in vivo, sequestration of serum TGFβ was assessed in A375 tumor-bearing NSG immune-reconstituted mice treated with either a-TGFβ or IgG-TGFβRII (5 mg kg⁻¹ per week) for 4 weeks (Supplementary Figure 2). At the endpoint, serum was collected from tail bleed and levels of TGFβ were detected using the TGFβ-1 Human ELISA Kit (ThermoFisher Scientific) following the manufacturer’s protocol.