Levels of TGFβ1 in mouse serum were measured using a Mouse TGF-beta 1 DuoSet ELISA kit (Catalog# DY1679-05 from R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s guidelines. Briefly, total TGFβ1 levels were measured in 40 μL of serum by mixing these with 10 μL 1 N HCl, followed by neutralization with 10 μL 1.2 N NaOH/0.5 M HEPES, according to the manufacturer’s sample activation procedure. These samples then were diluted 1:30 in assay diluent for measurement of total TGFβ1. ELISA plates were analyzed by reading absorbance using a microplate reader set to 450 nm with wavelength correction set to 570 nm.
Mouse tgf beta 1 duoset elisa kit
Manufactured by R&D Systems
Sourced in United States
The Mouse TGF-beta 1 DuoSet ELISA kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of mouse transforming growth factor beta 1 (TGF-beta 1) levels in cell culture supernatants, serum, and plasma.
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4 protocols using mouse tgf beta 1 duoset elisa kit
1
Quantifying Total TGF-β1 in Mouse Serum
2
Cytokine Quantification by ELISA
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Cytokine concentrations in the culture supernatants were determined using the Mouse TGF-beta 1 DuoSet ELISA kit (R&D Systems) or the Mouse TNF alpha, IL-6, or IL-12 ELISA Kits (Invitrogen). Values were quantified against a standard curve.
3
Quantifying Active TGFβ1 in Bone Marrow
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Active TGFb1 was assessed in bone marrow fluid using the Mouse TGF-beta 1 DuoSet ELISA Kit (R&D Systems) and DuoSet ELISA Ancillary Reagent Kit 1 (R&D Systems).
4
Quantification of Active TGFβ1 in Muscles
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Muscles were extracted at 7 dpi as described above and flash frozen in liquid nitrogen, then stored at –80°C. Tissues were thawed in ice-cold PBS, weighed, minced, and homogenized with 30 passes of a Dounce homogenizer in 500 uL of RIPA buffer (Thermo Fisher) with a protease inhibitor cocktail (Thermo Fisher). Total protein was quantified using a Pierce BCA Assay kit (Thermo Fisher) according to the manufacturer’s instructions. Active TGFβ1 was quantified using the mouse TGF beta 1 DuoSet ELISA kit (R&D Systems) according to the manufacturer’s instructions. Absorbances were measured on a Synergy Neo microplate reader.
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